title: Activity of heat shock genes' promoters in thermally contrasting animal species.
abstract: Heat shock gene promoters represent a highly conserved and universal system for the rapid induction of transcription after various stressful stimuli. We chose pairs of mammalian and insect species that significantly differ in their thermoresistance and constitutive levels of Hsp70 to compare hsp promoter strength under normal conditions and after heat shock (HS). The first pair includes the HSPA1 gene promoter of camel (Camelus dromedarius) and humans. It was demonstrated that the camel HSPA1A and HSPA1L promoters function normally in vitro in human cell cultures and exceed the strength of orthologous human promoters under basal conditions. We used the same in vitro assay for Drosophila melanogaster Schneider-2 (S2) cells to compare the activity of the hsp70 and hsp83 promoters of the second species pair represented by Diptera, i.e., Stratiomys singularior and D. melanogaster, which dramatically differ in thermoresistance and the pattern of Hsp70 accumulation. Promoter strength was also monitored in vivo in D. melanogaster strains transformed with constructs containing the S. singularior hsp70 ORF driven either by its own promoter or an orthologous promoter from the D. melanogaster hsp70Aa gene. Analysis revealed low S. singularior hsp70 promoter activity in vitro and in vivo under basal conditions and after HS in comparison with the endogenous promoter in D. melanogaster cells, which correlates with the absence of canonical GAGA elements in the promoters of the former species. Indeed, the insertion of GAGA elements into the S. singularior hsp70 regulatory region resulted in a dramatic increase in promoter activity in vitro but only modestly enhanced the promoter strength in the larvae of the transformed strains. In contrast with hsp70 promoters, hsp83 promoters from both of the studied Diptera species demonstrated high conservation and universality.
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia.
authors.forename: Lyubov N
authors.initials: LN
authors.lastname: Astakhova
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia.
authors.forename: Olga G
authors.initials: OG
authors.lastname: Zatsepina
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia.
authors.forename: Sergei Yu
authors.initials: SY
authors.lastname: Funikov
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia.
authors.forename: Elena S
authors.initials: ES
authors.lastname: Zelentsova
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia.
authors.forename: Natalia G
authors.initials: NG
authors.lastname: Schostak
authors.affiliation: Institute of Cytology and Genetics, The Siberian Branch of RAS, Prospekt Lavrentyeva 10,630090, Novosibirsk, Russia.
authors.forename: Konstantin E
authors.initials: KE
authors.lastname: Orishchenko
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia; Institute of Cell Biophysics RAS, Pushchino, Moscow region, 142290, Russia.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Evgen'ev
authors.affiliation: Engelhardt Institute of Molecular Biology RAS, Vavilov str. 32, Moscow, 119991, Russia.
authors.forename: David G
authors.initials: DG
authors.lastname: Garbuz
mesh_terms: D000818:Animals; D001483:Base Sequence; D002162:Camelus / Q000235:genetics; D002460:Cell Line; D004331:Drosophila melanogaster / Q000235:genetics; D017930:Genes, Reporter; D006360:Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D013045:Species Specificity; D016385:TATA Box; D015533:Transcriptional Activation
chemical_list: D006360:Heat-Shock Proteins; D049410:Luciferases, Renilla