Small heat shock protein Hsp67Bc plays a significant role in <i>Drosophila melanogaster</i> cold-stress tolerance

Malkeyeva, Dina; Kiseleva, Elena; Федорова, С. А.

doi:10.1242/jeb.219592

Cited by 12 publications

(10 citation statements)

References 39 publications

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“…Very few deregulated genes were shared between the uL11 mutants and corto (69 shared by uL11 K3A and corto , 29 shared by uL11 K3Y and corto , 14 shared by the three genotypes) ( Fig 8 ). We chose some genes to analyze their expression by RT-qPCR in the three mutants: CG13516 , one of the few genes deregulated in the three genotypes and the most down-regulated gene in uL11 K3Y , Hsp67Bc , encoding a small heat-shock protein involved in cold stress tolerance [ 40 ], which is up-regulated in corto L1 /corto 420 and uL11 K3Y , and GstE6 , encoding a Glutathione S-transferase up-regulated in uL11 K3A and uL11 K3Y . The RNA-seq data were confirmed by RT-qPCR for these three genes in the three mutants, except for Hsp67Bc for which the increase in corto L1 /corto 420 was not significant ( Fig 8 ).…”

Section: Resultsmentioning

confidence: 99%

Single amino-acid mutation in a Drosoph ila melanogaster ribosomal protein: An insight in uL11 transcriptional activity

et al. 2022

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The ribosomal protein uL11 is located at the basis of the ribosome P-stalk and plays a paramount role in translational efficiency. In addition, no mutant for uL11 is available suggesting that this gene is haplo-insufficient as many other Ribosomal Protein Genes (RPGs). We have previously shown that overexpression of Drosophila melanogaster uL11 enhances the transcription of many RPGs and Ribosomal Biogenesis genes (RiBis) suggesting that uL11 might globally regulate the level of translation through its transcriptional activity. Moreover, uL11 trimethylated on lysine 3 (uL11K3me3) interacts with the chromodomain of the Enhancer of Polycomb and Trithorax Corto, and both proteins co-localize with RNA Polymerase II at many sites on polytene chromosomes. These data have led to the hypothesis that the N-terminal end of uL11, and more particularly the trimethylation of lysine 3, supports the extra-ribosomal activity of uL11 in transcription. To address this question, we mutated the lysine 3 codon using a CRISPR/Cas9 strategy and obtained several lysine 3 mutants. We describe here the first mutants of D. melanogaster uL11. Unexpectedly, the uL11K3A mutant, in which the lysine 3 codon is replaced by an alanine, displays a genuine Minute phenotype known to be characteristic of RPG deletions (longer development, low fertility, high lethality, thin and short bristles) whereas the uL11K3Y mutant, in which the lysine 3 codon is replaced by a tyrosine, is unaffected. In agreement, the rate of translation decreases in uL11K3A but not in uL11K3Y. Co-immunoprecipitation experiments show that the interaction between uL11 and the Corto chromodomain is impaired by both mutations. However, Histone Association Assays indicate that the mutant proteins still bind chromatin. RNA-seq analyses from wing imaginal discs show that Corto represses RPG expression whereas very few genes are deregulated in uL11 mutants. We propose that Corto, by repressing RPG expression, ensures that all ribosomal proteins are present at the correct stoichiometry, and that uL11 fine-tunes its transcriptional regulation of RPGs.

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Section: Resultsmentioning

confidence: 99%

Single amino-acid mutation in a Drosoph ila melanogaster ribosomal protein: An insight in uL11 transcriptional activity

et al. 2022

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“…Very few deregulated genes were shared between the uL11 mutants and corto (69 shared by uL11 K3A and corto , 29 shared by uL11 K3Y and corto , 14 shared by the three genotypes) (Figure 7A). We chose some genes to analyze their expression by RT-qPCR in the three mutants: CG13516 , one of the few genes deregulated in the three genotypes and the most down-regulated gene in uL11 K3Y , Hsp67Bc, encoding a small heat-shock protein involved in cold stress tolerance (37), which is up-regulated in corto L1 /corto 420 and uL11 K3Y , and GstE6 , encoding a Glutathione S-transferase up-regulated in uL11 K3A and uL11 K3Y . The RNA-seq data were confirmed by RT-qPCR for these three genes in the three mutants, except for Hsp67Bc for which the increase in corto L1 /corto 420 was not significant (Figures 7B and C).…”

Section: Resultsmentioning

confidence: 99%

Single amino-acid mutation in a Drosophila melanogaster ribosomal protein: an insight in uL11 transcriptional activity

Grunchec

Deraze

Dardalhon-Cuménal

et al. 2021

Preprint

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The ribosomal protein uL11 is located at the basis of the ribosome P-stalk and plays a paramount role in translational efficiency. In addition, no mutant for uL11 is available suggesting that this gene is haplo-insufficient as many other Ribosomal Protein Genes (RPGs). We have previously shown that overexpression of Drosophila melanogaster uL11 enhances the transcription of many RPGs and Ribosomal Biogenesis genes (RiBis) suggesting that uL11 might globally regulate the level of translation through its transcriptional activity. Moreover, uL11 trimethylated on lysine 3 (uL11K3me3) interacts with the chromodomain of the Enhancer of Polycomb and Trithorax Corto, and both proteins co-localize with RNA Polymerase II at many sites on polytene chromosomes. These data have led to the hypothesis that the N-terminal end of uL11, and more particularly the trimethylation of lysine 3, supports the extra-ribosomal activity of uL11 in transcription. To address this question, we mutated the lysine 3 codon using a CRISPR/Cas9 strategy and obtained several lysine 3 mutants. We describe here the first mutants of D. melanogaster uL11. Unexpectedly, the uL11K3A mutant, in which the lysine 3 codon is replaced by an alanine, displays a genuine Minute phenotype known to be characteristic of RPG deletions (longer development, low fertility, high lethality, thin and short bristles) whereas the uL11K3Y mutant, in which the lysine 3 codon is replaced by a tyrosine, is unaffected. In agreement, the rate of translation decreases in uL11K3A but not in uL11K3Y. Co-immunoprecipitation experiments show that the interaction between uL11 and the Corto chromodomain is impaired by both mutations. RNA-seq analysis from wing imaginal discs shows enrichment in the GO categories glutathione metabolism for up-regulated genes in both uL11K3A and uL11K3Y mutants and regulation of transcription for down-regulated genes in uL11K3A only. Analysis of the cis-regulatory sequences of these genes suggests that uL11 might regulate transcription of target genes in concert with the couple of transcription factors Mad/Med that mediate response to the Bone Morphogenetic Protein (BMP) signaling pathway.

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“…Given the typical association with sHSPs and heat stress, we were somewhat surprised to find that HSP21, HSP24.6, HSP25.1, and HSP38 could each improve bacterial desiccation survival (Figure 2), but only HSP24.6 was sufficient to improve bacterial heat shock survival (Figure S4B). Notably, not all sHSPs function during heat shock, and some act as chaperones instead during cold stress or other conditions (66). It is possible that tardigrade sHSPs have, in fact, undergone selection for function during desiccation.…”

Section: Discussionmentioning

confidence: 99%

Tardigrade small heat shock proteins can limit desiccation-induced protein aggregation

Hibshman¹,

Carra

Goldstein

2022

Preprint

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Small heat shock proteins (sHSPs) are chaperones known for their response to heat stress. While sHSPs have well-characterized roles in heat tolerance, potential roles for sHSPs in desiccation tolerance have not been as thoroughly explored. We identified nine sHSPs from the genome of the tardigrade Hypsibius exemplaris, each containing a conserved alpha-crystallin domain flanked by disordered regions. Many of these sHSPs are highly expressed and in some cases are upregulated during desiccation. We found that tardigrade sHSPs were sufficient to improve desiccation tolerance when expressed in E. coli. Purification and subsequent analysis of two sHSPs, HSP21 and HSP24.6, revealed that these proteins can form large complexes in vitro, similar to oligomeric assemblies documented for other sHSPs. These proteins limited heat-induced aggregation of the model enzyme citrate synthase. Heterologous expression of HSP24.6 improved bacterial heat shock survival, and the protein significantly reduced heat-induced aggregation of soluble bacterial protein. Thus, HSP24.6 likely chaperones against protein aggregation to promote survival of heat stress. Furthermore, HSP21 and HSP24.6 also limited desiccation-induced aggregation and loss of function of citrate synthase. This suggests a mechanism by which tardigrade sHSPs promote desiccation tolerance: by limiting desiccation-induced protein aggregation, thereby maintaining proteostasis and supporting survival. These results suggest that sHSPs, classical chaperones, provide a mechanism of general stress resistance that can also be deployed to support survival during anhydrobiosis.

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Small heat shock protein Hsp67Bc plays a significant role in Drosophila melanogaster cold-stress tolerance

Cited by 12 publications

References 39 publications

Single amino-acid mutation in a Drosoph ila melanogaster ribosomal protein: An insight in uL11 transcriptional activity

Single amino-acid mutation in a Drosoph ila melanogaster ribosomal protein: An insight in uL11 transcriptional activity

Single amino-acid mutation in a Drosophila melanogaster ribosomal protein: an insight in uL11 transcriptional activity

Tardigrade small heat shock proteins can limit desiccation-induced protein aggregation

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