• Users Online: 875
  • Home
  • Print this page
  • Email this page
Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contacts Login 

Table of Contents
Year : 2022  |  Volume : 59  |  Issue : 2  |  Page : 145-153

Gamma radiation induced changes in expression of heat shock proteins (Hsc70 and Hsp83) in the dengue vector Aedes aegypti (L.)

1 Centre for Applied Genetics, J. B. Campus, Bangalore University, Bengaluru, India; Department of Entomology, Texas A&M University, College Station, Texas, USA
2 Centre for Applied Genetics, J. B. Campus, Bangalore University, Bengaluru, India
3 Environmental Assessment Division, Bhabha Atomic Research Centre, Mumbai, India
4 Radiation Biology and Health Science Division (BRNS-DAE), Bhabha Atomic Research Centre, Mumbai, India

Date of Submission11-Oct-2021
Date of Acceptance26-Nov-2021
Date of Web Publication08-Sep-2022

Correspondence Address:
N J Shetty
Centre for Applied Genetics, J. B. Campus, Bangalore University, Bengaluru 560056
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0972-9062.335770

Rights and Permissions

We aimed to assess the effect of gamma radiation on the expression of heat shock proteins Hsc70 and Hsp83 in Aedes aegypti. Adult males were irradiated with 50Gy of gamma radiation, and changes in the expression of proteins in SDS-PAGE gel bands corresponding to molecular weights ~60–75kDa and ~80–95kDa were analyzed at two different time points 6 and 12-hour post-irradiation, using a temporal mass spectrometry based semi-quantitative analysis. A 2-3-fold increase was observed in both proteins Hsc70 and Hsp83, at both time points. In addition, the experiment also revealed the overexpression of several other molecules such as Arginine Kinase - known to be upregulated in certain insects during stress, Esterase B1- implicated in insecticide resistance, and also down-regulation of the 26S proteasome non-ATPase regulatory subunit 1 and ubiquitin-activating enzyme E1 - both known to be involved in ubiquitin-mediated protein degradation. The results taken together with existing data on Hsp83 and Hsc70, indicate that these proteins may enhance the survival of Ae. aegypti following gamma radiation and could serve as molecular markers for the detection of radiation-induced stress.

Keywords: gamma radiation; heat shock proteins; Hsc70; Hsp83; Aedes aegypti

How to cite this article:
Shetty V, Shetty N J, Jha S K, Chaubey R C. Gamma radiation induced changes in expression of heat shock proteins (Hsc70 and Hsp83) in the dengue vector Aedes aegypti (L.). J Vector Borne Dis 2022;59:145-53

How to cite this URL:
Shetty V, Shetty N J, Jha S K, Chaubey R C. Gamma radiation induced changes in expression of heat shock proteins (Hsc70 and Hsp83) in the dengue vector Aedes aegypti (L.). J Vector Borne Dis [serial online] 2022 [cited 2022 Nov 28];59:145-53. Available from: https://www.jvbd.org/text.asp?2022/59/2/145/335770

  Introduction Top

Aedes aegypti is a well-known insect vector responsible for transmitting the viruses that cause dengue fever and the more severe dengue hemorrhagic fever[1]. Numerous techniques are being employed to restrict its growth and spread. One among them is the Sterile Insect Technique (SIT), a new-age approach that involves the generation of sterile males to check insect populations, usually using gamma radiation, and has been successfully applied on several insect species, including mosquito vectors[2],[3],[4].

Intentional irradiation aside, there is also increasing evidence to suggest that gamma radiation is an environmental stress factor that impacts all living organisms ubiquitously[5]. Insects perceive stress signals and respond with diverse strategies to defend against the stress, one of which is the regulation of protein expression[6]. Studies have confirmed that gamma radiation exposure can increase the levels of oxidative stress in insects, disturb the protein’s functional activity, and intensify the activity of protein oxidation processes[7],[8],[9]. There are a considerable number of studies investigating the effects of gamma radiation on Aedes, including its ability to induce sterility. Radiation-induced male sterility has been demonstrated in many mosquito species, including Ae. aegypti[10],[11],[12]. There are, however, very few reports of the molecular changes induced by gamma radiation that contribute to sterility.

Heat shock proteins (HSPs) are commonly studied in the context of insect stress management[13],[14], and gamma irradiation has been known to induce the activity of HSPs in different organisms[15],[16]. In vertebrates for instance, various radiation-responsive HSP genes have been identified from cells, tissues or individuals, and their roles have been characterized at the cellular and molecular levels[17],[18],[19]. The main role of HSPs is well known as molecular chaperones that promote proper assembly of denatured proteins, and hsp genes respond uniquely to different kinds of stress[13],[20]. It has been reported that HSPs play an important role in temperature stress-induced male sterility, male gametogenesis, and embryonic development in several drosophila species[21],[22],[23]. The HSPs in Ae. aegypti AeaHsp70 and AeaHsp83 have been proved to be important markers of temperature-induced stress. They are postulated to function as critical proteins to protect and enhance the survival of Ae. aegypti larvae and pupae subjected to temperature stress[24],[25]. Heat shock cognate 70 (Hsc70), an essential member of the HSP70 family, is an important stress-resistance protein found in insects and is known to protect them from environmental stresses[26]. The Heat Shock Proteins (HSPs), especially Hsp83/90 and Hsc70, are abundantly expressed in insects and are known modulators of insect survival[9],[27],[28]. Although, the practical application of measuring hsp gene products may be limited because their levels are altered by various external stimuli[13],[14],[29],[30], it has been suggested that they can serve as biomarkers of environmental contaminants, such as pesticides and metallic pollutants[31],[32]. There is however no mention in literature of HSPs in the context of gamma radiation induced stress in Aedes.

To this end, the current study was designed to determine the gamma radiation induced expression of heat shock proteins (Hsc70 and Hsp83) in Ae. aegypti. A dose equivalent to 50Gy that produced above 90% of sterility in the total irradiated males were selected for this study[33]. An MS-MS based proteomic approach was chosen, because it would allow the detection of unique peptides leading to a more specific identification of the HSPs induced. Further, this approach would allow the identification of few other proteins that are also possibly regulated in response to gamma radiation induced stress.

  Material & Methods Top

Mosquito rearing

Aedes aegypti larvae collected from the J.P Nagar area of Bengaluru, India were reared at 25 ± 10C and 75 ± 5% relative humidity under 14-hour photoperiod in the insectary of the Centre for Applied Genetics, Bangalore University, India following standard protocol[34].

Gamma irradiation experiments

Experiments were performed in three replicates along with a control. Overall, a total of 120 adult males, 2–3 days of age, were irradiated with 50Gy of gamma radiation from a60Co (Theratron 780-C machine) source with a dose rate of 253.56cGy/min, at the Kidwai Memorial Institute of Oncology, Bengaluru, India. The mosquitoes were placed in plastic boxes (5 X 4 X 2.5 cm) covered with fine net cap during irradiation. A dosimetry was used to quantify the dose received by the irradiated insects and confirm that all the doses delivered lay within a 5% error range. After irradiation, the mosquitoes were reared as described until further analysis. Another set of non-irradiated 60 adult males were maintained as controls (20 males in triplicates).

Preparation of crude homogenates

Whole-body homogenates were prepared by pooling twenty irradiated males each at two different time intervals, i.e., 6- and 12-hour post irradiation. A control set was prepared in a similar manner. The homogenized pellet was mixed with 500μl ice cold extraction buffer (50mM KPO4 buffer at pH 7.4), followed by centrifugation at 16,300g at 40C for 15 min. The clear supernatant was transferred to a new eppendorf tube and kept at -800C for approximately one week until analysis. Total protein concentration was determined by Bradford protein assay, using bovine serum albumin as the standard[35].

Sodium Dodecyl Sulphate (SDS) Polyacrylamide Gel Electrophoresis (PAGE)

Fifteen micrograms of protein from each sample (both experimental and control), were loaded onto a 10% SDS gel along with a standard protein maker ranging 10–250kD (Precision Plus Protein Standards (Dual Colour), Bio-Rad), and was run as described by Laemmli[36]. A total of three such gels were prepared to serve as technical replicates.

The gels were stained using colloidal Coomassie blue stain 250 (Bio-Rad), de-stained to remove excess stain, and then scanned, using a densitometric scanner to determine the concentration and molecular weight of each characteristic band relative to the protein molecular weight marker.

In-gel digestion and LC-MS/MS analysis

In-gel digestion, tryptic peptide extractions, and Nano-LC-MS/MS were conducted by the mass spectrometry facility, Centre for Cellular and Molecular Platforms (CCAMP), NCBS, Bengaluru, India following the methods of Shevchenko et al.[37].

The three SDS gel slabs were rinsed with water for a few hours, and the bands of interest labelled as A, B, C, D, E and F [Figure 1], were excised with a clean scalpel. Each band was cut into cubes (~1cubic millimetre) and transferred into labelled micro centrifuge tubes. Gel fragments from each tube were subjected to in gel digestion using Trypsin (Promega, Madison, WI) for approximately 12 h at 37°C. The digested peptides were reconstituted in 15μL of the 0.1% formic acid and 1μL of the same was injected on column and subjected to a 70 min RPLC gradient, followed by analysis on LTQ-Orbitrap-MS. The identity of the generated data was searched using MASCOT as the search engine against Swiss-prot, A. aegypti and TrEMBL databases.
Figure 1: A representative image of the Sodium Dodecyl Sulphate (SDS) Poly Acrylamide Gel Electrophoresis (PAGE) for proteins of post-irradiated adult males (at 6h and 12h time intervals) and control group of Aedes aegypti. The bands excised at each time point corresponding to molecular weight ranges ~60-75 and ~80-95 (kDa) are marked with letters A, B, C, D, E and F.

Click here to view

Data analysis

Progenesis LC-MS software was used for ion intensity-based label-free quantification and data analysis. Raw-files were imported and all replicate runs were aligned and normalized by the software to generate fold changes. Differentially expressed proteins exhibiting a fold change greater than 2-fold between the irradiated samples at two different time intervals (at 6- and 12-hr post-irradiation) and the controls were considered for analysis. Only those proteins identified at a false discovery rate of 1% or less were included. The unknown proteins were searched against nr database. The analyses were exported into an Excel spreadsheet containing the identified protein list including their corresponding scores, normalized fold change values (relative protein expression), ANOVA p-value (reliability of the measured differences are calculated between each group, are calculated using summed peptide ion abundances) and number of peptides matched to each protein.

Ethical statement: Not applicable

  Results & Discussion Top

This study is an account of a mass spectrometry-based relative quantification of proteins from SDS-PAGE bands of proteins extracted from adult male Ae. aegypti whole body lysate at 6 and 12 hours post gamma irradiation, corresponding to the ~60–75kDa and ~80–95kDa, respectively [Figure 1]. The analysis revealed a significant difference (p<0.05) in the relative abundance of a total of 21 proteins identified by MASCOT when the 6- and 12-hour post-irradiation samples were compared to the control. The results with the more than two (>2) fold change values derived from the Progenesis-based analysis are captured in [Table 1] & [Table 2]. Hsc70 and Hsp83 were both found to be upregulated in Ae. aegypti upon gamma irradiation. Interestingly, the study also revealed an up-regulation of enzymes such as Arginine kinase known to be upregulated in dipterans during stress, Esterase B1 implicated in insecticide resistance and downregulation of the 26S proteasome non-ATPase regulatory subunit 1 and ubiquitin-activating enzyme E1 that are involved in ubiquitin-mediated protein degradation.
Table 1: Differential expression profiles of selected proteins (MW ranges ~ 60–75kDa and ~80–95kDa) at 6h post irradiated Ae. aegypti

Click here to view
Table 2: Differential expression profiles of selected proteins (MW ranges ~ 60–75kDa and ~80–95kDa) at 12h post irradiated Ae. aegypti

Click here to view

The response of cells or organisms to heat shock and other stressors is connected to the induction of heat shock proteins (HSPs). In addition to their roles in protecting cells from stress, almost all HSPs are constitutively expressed in organisms at normal conditions, where they function as chaperones to ensure the correct folding of proteins and assist the translocation of proteins across intracellular membranes[38],[39],[40]. While there have been studies correlating the expression of heat shock protein to temperature shock, there are no reports of the HSP response to gamma irradiation in Aedes.

Beyond its role as molecular chaperones, some members of the HSP family have been reported to be involved in the development of sterility. Hsp70, for instance, plays a crucial role in spermatogenesis and thus has been linked to male sterility in Drosophila[41],[42]. Hsp70 family of the proteins located within the cytoplasm, one of the members of this family, Hsc70 constitutively expressed in spermatogonial cells[43]. It was also noted that Hsc70 is required as members of a chaperone complex for activating the ecdysone receptor which controls the successive stages of the insect’s life cycle and also contributes to other processes such as reproduction[44],[45]. Likewise, Huang et al.[28] found an inverse correlation between thermal protection and fecundity with respect to the overexpression of Hsp70 in Liriomyza huidobrensis. Temperature-induced male sterility has been reported in Drosophila buzzatii as well[23]. Also, the involvement of Hsp83 in spermatogenesis has been recorded[46].

Although gamma radiation based sterility has been extensively studied in Ae. aegypti particularly because of work on SIT[47],[48],[49], the plausible role of HSPs in the pathogenesis of sterility has not been looked at into. An earlier study from our lab recorded above 90% male sterility to 50Gy of gamma radiation exposure in Ae. aegypti[33]. Thus further investigation is necessary to validate the possible secondary role of HSPs in the induction of radiation-induced male sterility in the said species.

Hsp83 identified in the bands A and C corresponding to the ~80–95kDa bands of samples collected at 6- and 12-hours post-irradiation, respectively, was significantly up-regulated (~3 fold) at both time points, when compared to the control (band E) [Figure 1]. RNA expression studies have revealed a 40-to-50-fold increase in Hsp83 levels in Ae. aegypti larvae and pupae post heat shock treatment to 42oC[25]. Of course, the stress response with respect to gamma radiation and temperature cannot be compared in this case because there is a known lack of absolute correlation between mRNA and protein expression levels[50]. AeaHsp83 are important markers of stress and have been postulated to function as critical proteins involved in the protection and enhanced survival of Ae. aegypti[25]. A ~3- fold increase in the expression levels of this protein at 50 Gy exposures could explain the survivability of adults exposed to high doses of gamma radiation. Also, this increase in AeaHsp83 expression may be associated with the sterility that is induced at this dose[33].

Hsp genes in the same organism are likely to respond differently to different stimuli[16]. A heat shock study in Ae. aegypti, for instance, has shown that exposure to higher temperature has little effect on the RNA expression of AeaHsc70[25]. In our study, however, the analysis of gel bands B and D identified Hsc70 (Heat shock cognate 70) to be significantly (P<0.05) more abundant post-exposure to the gamma radiation. An approximate 2 and 2.5 fold up-regulation were observed following 6 and 12 hours of irradiation, respectively, when compared to control (band F). Although no studies on gamma-radiation induced HSP genes in mosquitoes are documented, there are a few studies on other insects. A study of the Indian meal moth, Plodia interpunctella, for instance, has also reported an increased level of Hsc70 in response gamma radiation exposure[16]. Hsc70 expression appears to be triggered by gamma radiation and not temperature stress in Ae. aegypti. However, the gamma radiation induced expression of Hsc70 seems to be conserved across various species of insects. Another study has shown that Chironomus ramosus larvae expressed Hsp70 upon gamma-radiation exposure and has postulated Hsp70 might be one of the gamma radiation-induced stress proteins required during the early stages of radiation stress management. Hsc70 thus does play a role in the tolerance of high doses of radiation induced stress in Ae. aegypti and hence could be considered as a potential biomarker for the detection of high doses of radiation induced stress.

The results of this study become more interesting in the light of the fact that several families of heat shock proteins known to be expressed in mosquitoes may have a cumulative role in determining the susceptibility to the virus[51]. In Anopheles gambiae, it has been shown that the expression of Hsc70B is induced not only by heat shock but also during an arbovirus infection, and Hsc70B protein is expressed to cope with cellular stress imposed during infection[52]. On the other hand, it was also shown that the Hsc70B protein product has important roles in homeostasis and suppression of o’yong-nyong virus replication in the vector, An. gambiae[53]. In this context, a 2-3 fold up regulation of Hsc70 and Hsp83 would thus indicate that at this dose, it may contribute to the susceptibility of Ae. aegypti to viruses.

Apart from the HSPs, the experiment also picked up few other proteins whose expressions were dysregulated upon irradiation. 19 other differentially expressed proteins, which have important biological functions, were identified, providing a basis for understanding their possible roles in response to gamma radiation stress. Of these, 13 proteins were more abundant when compared to that from control, while the remaining 6 proteins were less abundant [Table 1] & [Table 2]. The differentially expressed proteins identified from this study were ATP citrate synthase isoform X1, activity of metabolic enzyme pyruvate carboxylase, metabolic enzyme pyruvate kinase, glutamate semialdehyde dehydrogenase, tropomyosin invertebrate, protease m1 zinc metalloprotease, alanyl tRNA synthetase (exhibiting the highest fold change among all identified), arginine kinase, dihydropyrimidine dehydrogenase, Pyrroline 5 carboxylate dehydrogenase, Enzyme esterase B1, Nucleoside diphosphate kinase, and Putative calreticulin. The activities of proteins were low until 6-hour period and showed a gradual increase after 12-hour post irradiation over the control group. Although these findings are beyond the scope of our study, we feel it is important to discuss a few of them which are particularly relevant in the context of insect stress management.

Arginine kinase, which plays a role in the regulation of energy metabolism in cells, by catalyzing the formation of ATP from ADP (or vice versa) showed a 2.80-fold increase at 6 hours post radiation treatment. This enzyme is known to be prevalent in systems with fluctuating energy demands, acting as an energy buffering system[54] and an energy shuttle, delivering ATP generated by mitochondria to high energy requiring processes, such as membrane turnover[55]. Its presence is crucial when emerging from a dormant state and oxidative stress, acting as a modulator of energetic reserves under such conditions[56],[57].

Increased expression of arginine kinase, has been reported in other Dipterans exposed to infection[58],[59],[60]. It seemed to modulate the adaption of insects to adverse environments[61],[62]. Hence an upregulation of this enzyme post gamma radiation may play an important role in the survival of Ae. aegypti.

A 2.38-fold upregulation was observed in another enzyme Esterase B1 in Ae. aegypti at 12-hours post-irradiation. Interestingly, elevated levels of Esterase B1 have been observed in several insecticide resistant strains of Ae. aegypti, possibly to overcome insecticide induced stress[63],[64]. An overproduction of esterase B1 has also been recorded in insecticide resistance strains of few species of Culex against organophosphates[65]. An increased activity of esterase was noted in a pyrethroid resistant population of Anopheles albimanus Wiedemann. If esterase levels are an indication of insecticide resistance, then this implies that gamma radiation could influence the susceptibility of Ae. aegypti to insecticides. Although, of course, this would warrant more temporal analyses to conclude the expression levels of the enzyme beyond the 12 hours’ time point before such a correlation is drawn.

Two proteins required for embryonic, larval and germline development[66],[67] 26S proteasome non-ATPase regulatory subunit 1 and ubiquitin activating enzyme E1, were found to be downregulated in our dataset. The ubiquitin-proteasome system, through influencing protein stability plays an essential role in cell cycle regulation, stress, DNA repair, and carcinogenesis[68],[69],[70]. The polyubiquitinated proteins generated by ubiquitin-conjugating system serve as substrates for the proteasome[71],[72]. In short, proteins modified through one of the ubiquitin-activating enzyme E1are further targeted by the 26S proteasome for degradation[73]. The results of this study are therefore in agreement with a previous study that reported that oxidative stress induces the loss of activity of 26S proteosome[74].

  Conclusion Top

Response patterns of Ae. aegypti adults to gamma radiation stress conditions are complex, as the differentially abundant proteins are involved in multiple functional categories. The present study has identified a number of differentially expressed proteins, which can be chosen for further validation, and understanding the specific role of these proteins will provide new insights into the adaptive mechanisms of Ae. aegypti in response to gamma radiation stress. Our experiments revealed that gamma irradiation influenced the expression profile of HSP genes in Ae. aegypti. Higher expressions of these proteins suggest that they could be involved in the enhancement of the survival of Ae. aegypti and thus might be a potentially useful tool as a molecular marker for detection of radiation-induced stress.

Conflict of interest: None

  References Top

Ahmad I, S Astari, M Tan. Resistance of Aedes aegypti (Diptera: Culicidae) in 2006 to pyrethroid insecticides in Indonesia and its association with oxidase and esterase levels. Pakistan Journal of Biological Sciences 2007; 10(20): 3688–92.  Back to cited text no. 1
Knipling EF. Sterile Technique – Principles involved, Current Application, Limitations, and Future Application. Genetics of Insect Vectors of Disease, R.P. JW Wright, Editor, Elsevier: Amsterdam 1967; 587.  Back to cited text no. 2
Reinhardt K. Reproductive potential of gamma-irradiated males of the meadow grasshopper Chorthippus parallelus (Zetterstedt) (Orth., Acrididae). Journal of Applied Entomology 1999; 123(9): 519–523.  Back to cited text no. 3
Robinson AS. Mutations and their use in insect control. Mutatation Research 2002; 511(2): 113–32.  Back to cited text no. 4
Zhikrevetskaya S. Effect of Low Doses (5-40 cGy) of Gamma-irradiation on Lifespan and Stress-related Genes Expression Profile in Drosophila melanogaster. PLoS One 2015; 10(8): e0133840.  Back to cited text no. 5
Meng JY. Ultraviolet light-induced oxidative stress: effects on antioxidant response of Helicoverpa armigera adults. Journal of Insect Physiology 2009; 55(6): 588–92.  Back to cited text no. 6
Parashar V. The effects of age on radiation resistance and oxidative stress in adult Drosophila melanogaster. Radiation Research 2008; 169(6): 707-11.  Back to cited text no. 7
Datkhile KD. Increased level of superoxide dismutase (SOD) activity in larvae of Chironomus ramosus (Diptera: Chironomidae) subjected to ionizing radiation. Comparative Biochemistry and Physiology Part C. Toxicology & Pharmacology 2009; 149(4): 500–6.  Back to cited text no. 8
Zhao L, Jones W. Expression of heat shock protein genes in insect stress responses. Invertebrate Survival Journal 2012; 9: 93–101.  Back to cited text no. 9
Shetty NJ. Genetic control of mosquitoes: Chromosomal translocations and inherited semi-sterility in Culex quinquefasciatus - a filarial mosquito. Journal of Cytology and Genetics 1993; 28: 181–187.  Back to cited text no. 10
Helinski ME, AG Parker, BG Knols. Radiation-induced sterility for pupal and adult stages of the malaria mosquito Anopheles arabiensis. Malaria Journal 2006; 5: 41.  Back to cited text no. 11
Asman M, KS Rai. Developmental effects of ionizing radiation in Aedes aegypti. Journal of Medical Entomology 1972; 9(5): 468–78.  Back to cited text no. 12
Denlinger DL, JM Giebultowicz, DS Saunders. Preface, in Insect Timing: Circadian Rhythmicity to Seasonality. 2001, Elsevier Science B.V.: Amsterdam. p. v.  Back to cited text no. 13
Feder ME, GE Hofmann. Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annual Reveiw of Physiology 1999; 61: 243–82.  Back to cited text no. 14
Datkhile KD. Hsp70 expression in Chironomus ramosus exposed to gamma radiation. International Journal of Radiation Biology 2011; 87(2): 213–21.  Back to cited text no. 15
Shim JK. Gamma irradiation effects on the induction of three heat shock protein genes (piac25, hsc70 and hsp90) in the Indian meal moth, Plodia interpunctella. Journal of Stored Products Research 2009; 45(2): 75–81.  Back to cited text no. 16
Fornace AJ Jr. Mammalian genes induced by radiation; activation of genes associated with growth control. Annual Reveiw of Genetics 1992; 26: 507–26.  Back to cited text no. 17
Calini V, C Urani, M Camatini. Overexpression of HSP70 is induced by ionizing radiation in C3H 10T1/2 cells and protects from DNA damage. Toxicology In Vitro 2003; 17(5-6): 561–6.  Back to cited text no. 18
Lee HJ. Radioprotective effect of heat shock protein 25 on submandibular glands of rats. American Journal of Pathology 2006; 169(5): 1601–11.  Back to cited text no. 19
Lindquist S, EA Craig. The heat-shock proteins. Annual Reveiw of Genetics 1988; 22: 631–77.  Back to cited text no. 20
Haass C, U Klein, PM Kloetzel. Developmental expression of Drosophila melanogaster small heat-shock proteins. Journal of Cell Science 1990; 96: 413–8.  Back to cited text no. 21
Joanisse DR. Small heat shock proteins ofDrosophila: Developmental expression and functions. Journal of Biosciences 1998; 23(4): 369–376.  Back to cited text no. 22
Vollmer JH. Heat and cold-induced male sterility in Drosophila buzzatii: genetic variation among populations for the duration of sterility. Heredity 2004; 92(3): 257–62.  Back to cited text no. 23
Zhao L. Identification of genes differentially expressed during heat shock treatment in Aedes aegypti. Journal of Medical Entomology 2009; 46(3): 490–5.  Back to cited text no. 24
Zhao L. Expression of AeaHsp26 and AeaHsp83 in Aedes aegypti (Diptera: Culicidae) larvae and pupae in response to heat shock stress. Journal of Medical Entomology 2010; 47(3): 367–75.  Back to cited text no. 25
Sun Y. Identification of heat shock cognate protein 70 gene (Alhsc70) of Apolygus lucorum and its expression in response to different temperature and pesticide stresses. Insect Science 2016; 23(1): 37–49.  Back to cited text no. 26
Hong SM. Efficient soluble protein production on transgenic silkworms expressing cytoplasmic chaperones. Applied Microbiology and Biotechnology 2010; 87(6): 2147–56.  Back to cited text no. 27
Huang LH, B Chen, L Kang. Impact of mild temperature hardening on thermotolerance, fecundity, and Hsp gene expression in Liriomyza huidobrensis. Journal of Insect Physiology 2007; 53(12): 1199–205.  Back to cited text no. 28
Mahroof R. Expression patterns of three heat shock protein 70 genes among developmental stages of the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae). Comparative Biochemistry and Physiology Part A. Molecular & Integrative Physiology 2005; 141(2): 247–56.  Back to cited text no. 29
Huang LH, L Kang. Cloning and interspecific altered expression of heat shock protein genes in two leafminer species in response to thermal stress. Insect Molecular Biology 2007; 16(4): 491–500.  Back to cited text no. 30
Sanders BM. Stress proteins in aquatic organisms: an environmental perspective. Critical Reviews in Toxicology 1993; 23(1): 49–75.  Back to cited text no. 31
Lewis S, et al. Stress proteins (HSP’s): Methods of Detection and Their Use as an Environmental Biomarker. Ecotoxicology 1999; 8(5): 351–368.  Back to cited text no. 32
Shetty V. Effect of gamma radiation on life history traits of Aedes aegypti (L.). Parasite Epidemiology and Control 2016; 1(2): 26–35.  Back to cited text no. 33
Shetty NJ. Chromosomal translocations and semisterility in the malaria vector Anopheles fluviatilis James. Indian Journal of Malariology 1983; 20: 45–48.  Back to cited text no. 34
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 1976; 72: 248–54.  Back to cited text no. 35
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259): 680–5.  Back to cited text no. 36
Shevchenko A. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nature Protocol 2006; 1(6): 2856–60.  Back to cited text no. 37
Neupert W. Protein import into mitochondria. Annual Reveiw of Biochemistry 1997; 66: 863–917.  Back to cited text no. 38
Hartl FU, M Hayer-Hartl. Molecular chaperones in the cytosol: from nascent chain to folded protein. Science 2002; 295(5561): 1852–8.  Back to cited text no. 39
Nollen EAA & RI Morimoto. Chaperoning signaling pathways: molecular chaperones as stress-sensing ‘heat shock’ proteins. Journal of Cell Science 2002; 115(14): 2809–2816.  Back to cited text no. 40
Li K. Drosophila Centrosomin Protein is Required for Male Meiosis and Assembly of the Flagellar Axoneme. The Journal of Cell Biology 1998; 141(2): 455–467.  Back to cited text no. 41
Sorensen JG, et al. Expression of the heat-shock protein HSP70 in Drosophila buzzatii lines selected for thermal resistance. Hereditas 1999; 131(2): 155–64.  Back to cited text no. 42
Edward ME. HSP70 Chaperones in Spermatogenesis in The testis, E. Goldberg, Editor, Springer New York 2000; 133–142.  Back to cited text no. 43
Riddiford M. Hormones and Drosophila Development, in The Development of Drosophila melanogaster, A.M.M.a.A. Bate, Editor, Cold Spring Harbor: Cold Spring Harbor Laboratory Press 1993; 899–940.  Back to cited text no. 44
Arbeitman MN, DS Hogness. Molecular chaperones activate the Drosophila ecdysone receptor, an RXR heterodimer. Cell 2000; 101.  Back to cited text no. 45
Yue L. Genetic analysis of viable Hsp90 alleles reveals a critical role in Drosophila spermatogenesis. Genetics 1999; 151(3): 1065–79.  Back to cited text no. 46
Rai KS, PT McDonald, SM Asman. Cytogenetics of Two Radiation-Induced, Sex-Linked Translocations in the Yellow-Fever Mosquito, AEDES AEGYPTI. Genetics 1970; 66(4): 635–651.  Back to cited text no. 47
Rai KS, KK Grover, SG Suguna. Genetic manipulation of Aedes aegypti: incorporation and maintenance of a genetic marker and a chromosomal translocation in natural populations. Bulletin of the World Health Organization 1973; 48(1): 49–56.  Back to cited text no. 48
Rodriguez PH. Effects on the productivity of irradiated male populations of Aedes aegypti (Diptera: Culicidae). J Med Entomol 1977; 14(4): 493–4.  Back to cited text no. 49
Nie L, G Wu, W Zhang. Correlation between mRNA and protein abundance in Desulfovibrio vulgaris: a multiple regression to identify sources of variations. Biochemical and Biophysical Research Communications 2006; 339(2): 603–10.  Back to cited text no. 50
Yadav P. Effect of temperature and insecticide stresses on Aedes aegypti larvae and their influence on the susceptibility of mosquitoes to dengue-2 virus. Southeast Asian journal of tropical medicine and public health 2005; 36(5): 1139.  Back to cited text no. 51
Kang S. Ex vivo promoter analysis of antiviral heat shock cognate 70B gene in Anopheles gambiae. Virology Journal 2008; 5: 136.  Back to cited text no. 52
Sim C. Anopheles gambiae heat shock protein cognate 70B impedes o’nyong-nyong virus replication. BMC Genomics 2007; 8(1): 1–12.  Back to cited text no. 53
Canonaco F. Functional expression of phosphagen kinase systems confers resistance to transient stresses in Saccharomyces cerevisiae by buffering the ATP pool. Journal of Biological Chemistry 2002; 277(35): 31303–9.  Back to cited text no. 54
Kucharski R, R Maleszka. Arginine kinase is highly expressed in the compound eye of the honey bee, Apis mellifera. Gene 1998; 211(2): 343–9.  Back to cited text no. 55
Alonso GD. Arginine kinase of the flagellate protozoa Trypanosoma cruzi. Regulation of its expression and catalytic activity. FEBS Letters 2001; 498(1): 22–5.  Back to cited text no. 56
Miranda MR.Trypanosoma cruzi: Oxidative stress induces arginine kinase expression. Experimental Parasitology 2006; 114(4): 341–4.  Back to cited text no. 57
Levy F, P Bulet L. Ehret-Sabatier. Proteomic analysis of the systemic immune response of Drosophila. Molecular & Cellular Proteomics 2004; 3(2): 156–66.  Back to cited text no. 58
Vierstraete E. The instantly released Drosophila immune proteome is infection-specific. Biochemical and Biophysical Research Communications 2004; 317(4): 1052–60.  Back to cited text no. 59
Tchankouo-Nguetcheu S. Differential protein modulation in midguts of Aedes aegypti infected with chikungunya and dengue 2 viruses. PLoS One 2010; 5(10).  Back to cited text no. 60
Voncken F. The phosphoarginine energy-buffering system of trypanosoma brucei involves multiple arginine kinase isoforms with different subcellular locations. PLoS One 2013; 8(6).  Back to cited text no. 61
Pereira CA. Arginine kinase: a potential pharmacological target in trypanosomiasis. Infectious Disorders-Drug Targets 2014; 14(1): 30–6.  Back to cited text no. 62
Saavedra-Rodriguez K. Differential transcription profiles in Aedes aegypti detoxification genes after temephos selection. Insect Mol Biol 2014; 23(2): 199–215.  Back to cited text no. 63
Sousa Polezzi RdCs, HEMdC Bicudo. Effect of phenobarbital on inducing insecticide tolerance and esterase changes in Aedes aegypti (Diptera: Culicidae). Genetics and Molecular Biology 2004; 27: 275–283.  Back to cited text no. 64
Mouches, C. Overproduction of detoxifying esterases in organophosphate-resistant Culex mosquitoes and their presence in other insects. Proceedings of the National Academy of Sciences 1987; 84(8): 2113–6.  Back to cited text no. 65
Gonczy P. Functional genomic analysis of cell division in C. elegans using RNAi of genes on chromosome III. Nature 2000; 408(6810): 331–6.  Back to cited text no. 66
Takahashi M. Reverse genetic analysis of the Caenorhabditis elegans 26S proteasome subunits by RNA interference. Biological Chemistry 2002; 383(7-8): 1263–6.  Back to cited text no. 67
Choe KP, AJ Przybysz, K Strange. The WD40 repeat protein WDR-23 functions with the CUL4/DDB1 ubiquitin ligase to regulate nuclear abundance and activity of SKN-1 in Caenorhabditis elegans. Molecular Cell Biology 2009; 29(10): 2704–15.  Back to cited text no. 68
Chondrogianni N, ES Gonos. Structure and function of the ubiquitin-proteasome system: modulation of components. Progress in molecular biology and translational science 2012; 109: 41–74.  Back to cited text no. 69
Chondrogianni N. Proteasome activation delays aging in vitro and in vivo. Free Radical Biology and Medicine 2014; 71: 303–20.  Back to cited text no. 70
De la Cova C, I Greenwald. SEL-10/Fbw7-dependent negative feedback regulation of LIN-45/Braf signaling in C. elegans via a conserved phosphodegron. Genes & Development 2012; 26(22): 2524–35.  Back to cited text no. 71
De Strooper B. Proteases and proteolysis in Alzheimer disease: a multifactorial view on the disease process. Physiological Reviews 2010. 90(2): 465–94.  Back to cited text no. 72
Depuydt G. Reduced insulin/insulin-like growth factor-1 signaling and dietary restriction inhibit translation but preserve muscle mass in Caenorhabditis elegans. Molecular & Cellular Proteomics 2013; 12(12): 3624–39.  Back to cited text no. 73
Wang X. Regulation of the 26S proteasome complex during oxidative stress. Science Signaling 2010; 3(151): 88.  Back to cited text no. 74


  [Figure 1]

  [Table 1], [Table 2]


    Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
    Access Statistics
    Email Alert *
    Add to My List *
* Registration required (free)  

  Material & M...Results & Di...
  In this article
Article Figures
Article Tables

 Article Access Statistics
    PDF Downloaded93    
    Comments [Add]    

Recommend this journal