An intron-containing, heat-inducible stress-70 gene in the millipede Tachypodoiulus niger (Julidae, Diplopoda)
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The highly conserved part of the nucleotide-binding domain of the hsp70 gene family was amplified from the soil diplopod Tachypodoiulus niger (Julidae, Diplopoda). Genomic DNA yielded 701, 549 and 540 bp sequences, whereas cDNA from heat shocked animals produced only one distinct fragment of 543 bp. The sequences could be classified as a 70 kDa heat shock protein (hsp70), the corresponding 70 kDa heat shock cognate (hsc70) and a glucose-related hsp70 homologue (grp78). Comparisons of genomic and cDNA sequences of hsc70 identified two introns within the consensus sequence. Generally, stress-70 expression levels were low, which hampered successful RT-PCR and subsequent subcloning. Following experimental heat shock, however, the spliced hsc70 was amplified predominantly, instead of its inducible homologue hsp70. This finding suggests that microevolution in this soil-dwelling arthropod is directed towards low constitutive stress-70 levels and that the capacity for stress-70 induction presumably is limited. hsc70, albeit having introns, apparently is inducible and contributes to the stress-70 response.
KeywordsStress response Soil invertebrate Arthropods Heat shock proteins Heat shock cognates Glucose-related proteins Biomarker
We gratefully acknowledge the support by D. Ammermann, N.K. Jacob, M. Knigge, T. Monsinjon and R. Paxton. The authors are also grateful to M. Bulant for assistance with the phylogenetic analysis. This study received funding from the German Research Council (Grant No. Ko 1978/2-1/2).
- De Pomerai DI (1996) Heat schock proteins as biomarkers of pollution. Human and Exp. Toxicol 15:279–285Google Scholar
- Feder ME, Krebs RA (1998) Natural and genetic engeneering of the heat-shock protein hsp70 in Drosophila melanogaster: consequences for thermotolerance. Am Zool 38:503–517Google Scholar
- Jiao C, Wang Z, Li F, Zhang C, Xiang J (2004) Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 (HSC70) in Fenneropenaeus chinensis. Chin Sci Bull 49:2385–2393Google Scholar
- Sanders BM (1990) Stress proteins: potential as multitiered biomarkers. Lewis Publishers, CRC Press, Boca Raton, USAGoogle Scholar
- Sorger P (1991) Heat shock factor and the heat shock response. Cell 65:363–366Google Scholar
- Zanger M, Alberti G, Kuhn M, Köhler H-R (1996) The stress −70 protein family in diplopods: induction and characterization. J. Comp. Physiol. B 156:622–627Google Scholar
- Zhang D-Z, Hewitt GM (1998) Special DNA extraction methods for some animal species. Chapman & Hall, London, UKGoogle Scholar
- Zhang G, Fang X, Guo X, Li L, Luo R, Xu F et al. (2012) The oyster genome reveals stress adaptation and complexity of shell formation. Nature 490(7418):49–54 Google Scholar