Abstract
The transcription factor heat shock factor-1 (HSF-1) regulates the heat shock response (HSR), a cytoprotective response induced by proteotoxic stresses. Data from model organisms has shown that HSF-1 also has non-stress biological roles, including roles in the regulation of development and longevity. To better study HSF-1 function, we created a C. elegans strain containing HSF-1 tagged with GFP at its endogenous locus utilizing CRISPR/Cas9-guided transgenesis. We show that the HSF-1::GFP CRISPR worm strain behaves similarly to wildtype worms in response to heat and other stresses, and in other physiological processes. HSF-1 was expressed in all tissues assayed. Immediately following the initiation of reproduction, HSF-1 formed nuclear stress bodies, a hallmark of activation, throughout the germline. Upon the transition to adulthood, of HSF-1 nuclear stress bodies appeared in most somatic cells. Genetic loss of the germline suppressed nuclear stress body formation with age, suggesting that the germline influences HSF-1 activity. Interestingly, we found that various neurons did not form nuclear stress bodies after transitioning to adulthood. Therefore, the formation of HSF-1 nuclear stress bodies upon the transition to adulthood does not occur in a synchronous manner in all cell types. In sum, these studies enhance our knowledge of the expression and activity of the aging and proteostasis factor HSF-1 in a tissue-specific manner with age.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
References
Ahn SG, Thiele DJ (2003) Redox regulation of mammalian heat shock factor 1 is essential for Hsp gene activation and protection from stress. Genes Dev 17(4):516–528
Arantes-Oliveira N, Apfeld J, Dillin A, Kenyon C (2002) Regulation of life-span by germ-line stem cells in Caenorhabditis elegans. Science 295(5554):502–505
Ben-Zvi A, Miller EA, Morimoto RI (2009) Collapse of proteostasis represents an early molecular event in Caenorhabditis elegans aging. Proc Natl Acad Sci 106(35):14914–14919
Berman JR, Kenyon C (2006) Germ-cell loss extends C. elegans life span through regulation of DAF-16 by kri-1 and lipophilic-hormone signaling. Cell 124(5):1055–1068
Biamonti G (2004) Nuclear stress bodies: a heterochromatin affair? Nat Rev Mol Cell Biol 5(6):493–498. https://doi.org/10.1038/nrm1405
Biamonti G, Vourc’h C (2010) Nuclear stress bodies. Cold Spring Harb Perspect Biol 2(6):a000695. https://doi.org/10.1101/cshperspect.a000695
Bohnert KA, Kenyon C (2017) A lysosomal switch triggers proteostasis renewal in the immortal C. elegans germ lineage. Nature 551(7682):629
Brunquell J, Morris S, Lu Y, Cheng F, Westerheide SD (2016) The genome-wide role of HSF-1 in the regulation of gene expression in Caenorhabditis elegans. BMC Genomics 17:559. https://doi.org/10.1186/s12864-016-2837-5
Burkewitz K, Choe K, Strange K (2011) Hypertonic stress induces rapid and widespread protein damage in C. elegans. Am J Physiol Cell Physiol 301(3):C566–C576. https://doi.org/10.1152/ajpcell.00030.2011
Chalfie M, Sulston J (1981) Developmental genetics of the mechanosensory neurons of Caenorhabditis elegans. Dev Biol 82(2):358–370. https://doi.org/10.1016/0012-1606(81)90459-0
Chiang WC, Ching TT, Lee HC, Mousigian C, Hsu AL (2012) HSF-1 regulators DDL-1/2 link insulin-like signaling to heat-shock responses and modulation of longevity. Cell 148(1-2):322–334. https://doi.org/10.1016/j.cell.2011.12.019
Das S, Ooi FK, Cruz Corchado J, Fuller LC, Weiner JA, Prahlad V (2020) Serotonin signaling by maternal neurons upon stress ensures progeny survival. Elife 9. https://doi.org/10.7554/eLife.55246
Denegri M, Moralli D, Rocchi M, Biggiogera M, Raimondi E, Cobianchi F, de Carli L, Riva S, Biamonti G (2002) Human chromosomes 9, 12, and 15 contain the nucleation sites of stress-induced nuclear bodies. Mol Biol Cell 13(6):2069–2079. https://doi.org/10.1091/mbc.01-12-0569
Dickinson DJ, Ward JD, Reiner DJ, Goldstein B (2013) Engineering the Caenorhabditis elegans genome using Cas9-triggered homologous recombination. Nat Methods 10(10):1028–1034
Duchateau A, de Thonel A, El Fatimy R, Dubreuil V, Mezger V (2020) The “HSF connection”: pleiotropic regulation and activities of Heat Shock Factors shape pathophysiological brain development. Neurosci Lett 725:134895. https://doi.org/10.1016/j.neulet.2020.134895
Emtage L, Gu G, Hartwieg E, Chalfie M (2004) Extracellular proteins organize the mechanosensory channel complex in C. elegans touch receptor neurons. Neuron 44(5):795–807. https://doi.org/10.1016/j.neuron.2004.11.010
Erkekoglu P, Baydar T (2014) Acrylamide neurotoxicity. Nutr Neurosci 17(2):49–57. https://doi.org/10.1179/1476830513Y.0000000065
Eymery A, Souchier C, Vourc’h C, Jolly C (2010) Heat shock factor 1 binds to and transcribes satellite II and III sequences at several pericentromeric regions in heat-shocked cells. Exp Cell Res 316(11):1845–1855. https://doi.org/10.1016/j.yexcr.2010.02.002
Gaglia G, Rashid R, Yapp C, Joshi GN, Li CG, Lindquist SL, Sarosiek KA, Whitesell L, Sorger PK, Santagata S (2020) HSF1 phase transition mediates stress adaptation and cell fate decisions. Nat Cell Biol 22(2):151–158. https://doi.org/10.1038/s41556-019-0458-3
Giordano M, Infantino L, Biggiogera M, Montecucco A, Biamonti G (2020) Heat shock affects mitotic segregation of human chromosomes bound to stress-induced satellite III RNAs. Int J Mol Sci 21(8). https://doi.org/10.3390/ijms21082812
Gomez-Pastor R, Burchfiel ET, Thiele DJ (2017) Regulation of heat shock transcription factors and their roles in physiology and disease. Nat Rev Mol Cell Biol 19:4–19. https://doi.org/10.1038/nrm.2017.73
Hartl FU, Bracher A, Hayer-Hartl M (2011) Molecular chaperones in protein folding and proteostasis. Nature 475(7356):324–332. https://doi.org/10.1038/nature10317
Hsin H, Kenyon C (1999) Signals from the reproductive system regulate the lifespan of C. elegans. Nature 399(6734):362–366
Hsu A-L, Murphy CT, Kenyon C (2003) Regulation of aging and age-related disease by DAF-16 and heat-shock factor. Science 300(5622):1142–1145
Jolly C, Morimoto R, Robert-Nicoud M, Vourc’h C (1997) HSF1 transcription factor concentrates in nuclear foci during heat shock: relationship with transcription sites. J Cell Sci 110(Pt 23):2935–2941
Jolly C, Usson Y, Morimoto RI (1999) Rapid and reversible relocalization of heat shock factor 1 within seconds to nuclear stress granules. Proc Natl Acad Sci U S A 96(12):6769–6774
Jolly C, Konecny L, Grady DL, Kutskova YA, Cotto JJ, Morimoto RI, Vourc’h C (2002) In vivo binding of active heat shock transcription factor 1 to human chromosome 9 heterochromatin during stress. J Cell Biol 156(5):775–781
Jolly C, Metz A, Govin J, Vigneron M, Turner BM, Khochbin S, Vourc’h C (2004) Stress-induced transcription of satellite III repeats. J Cell Biol 164(1):25–33
Joutsen J, Sistonen L (2019) Tailoring of proteostasis networks with heat shock factors. Cold Spring Harb Perspect Biol 11(4). https://doi.org/10.1101/cshperspect.a034066
Kamath RS, Fraser AG, Dong Y, Poulin G, Durbin R, Gotta M et al (2003) Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421(6920):231–237
Labbadia J, Morimoto RI (2015) Repression of the heat shock response is a programmed event at the onset of reproduction. Mol Cell 59(4):639–650
Li J, Chauve L, Phelps G, Brielmann RM, Morimoto RI (2016) E2F coregulates an essential HSF developmental program that is distinct from the heat-shock response. Genes Dev 30(18):2062–2075. https://doi.org/10.1101/gad.283317.116
Martinez-Balbas MA, Dey A, Rabindran SK, Ozato K, Wu C (1995) Displacement of sequence-specific transcription factors from mitotic chromatin. Cell 83(1):29–38. https://doi.org/10.1016/0092-8674(95)90231-7
Mercier PA, Winegarden NA, Westwood JT (1999) Human heat shock factor 1 is predominantly a nuclear protein before and after heat stress. J Cell Sci 112(Pt 16):2765–2774
Merritt C, Rasoloson D, Ko D, Seydoux G (2008) 3′ UTRs are the primary regulators of gene expression in the C. elegans germline. Curr Biol 18(19):1476–1482
Morley JF, Morimoto RI (2004) Regulation of longevity in Caenorhabditis elegans by heat shock factor and molecular chaperones. Mol Biol Cell 15(2):657–664. https://doi.org/10.1091/mbc.E03-07-0532
Morton EA, Lamitina T (2013) Caenorhabditis elegans HSF-1 is an essential nuclear protein that forms stress granule-like structures following heat shock. Aging Cell 12(1):112–120. https://doi.org/10.1111/acel.12024
Nussbaum-Krammer CI, Morimoto RI (2014) Caenorhabditis elegans as a model system for studying non-cell-autonomous mechanisms in protein-misfolding diseases. Dis Model Mech 7(1):31–39
Ooi FK, Prahlad V (2017) Olfactory experience primes the heat shock transcription factor HSF-1 to enhance the expression of molecular chaperones in C. elegans. Sci Signal 10(501):eaan4893. https://doi.org/10.1126/scisignal.aan4893
Prahlad V, Cornelius T, Morimoto RI (2008) Regulation of the cellular heat shock response in Caenorhabditis elegans by thermosensory neurons. Science sas320(5877):811–814. https://doi.org/10.1126/science.1156093
Priess JR, Schnabel H, Schnabel R (1987) The glp-1 locus and cellular interactions in early C. elegans embryos. Cell 51(4):601–611. https://doi.org/10.1016/0092-8674(87)90129-2
Shemesh N, Shai N, Ben-Zvi A (2013) Germline stem cell arrest inhibits the collapse of somatic proteostasis early in Caenorhabditis elegans adulthood. Aging Cell 12(5):814–822. https://doi.org/10.1111/acel.12110
Singh V, Aballay A (2006) Heat-shock transcription factor (HSF)-1 pathway required for Caenorhabditis elegans immunity. Proc Natl Acad Sci U S A 103(35):13092–13097. https://doi.org/10.1073/pnas.0604050103
Sulston JE, Hodgkin J (1988) Methods. In: Wood WB (ed) The Nematode Caenorhabditis elegans. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 587–606
Tatum MC, Ooi FK, Chikka MR, Chauve L, Martinez-Velazquez LA, Steinbusch HW et al (2015) Neuronal serotonin release triggers the heat shock response in C. elegans in the absence of temperature increase. Curr Biol 25(2):163–174. https://doi.org/10.1016/j.cub.2014.11.040
Trott A, West JD, Klaic L, Westerheide SD, Silverman RB, Morimoto RI, Morano KA (2008) Activation of heat shock and antioxidant responses by the natural product celastrol: transcriptional signatures of a thiol-targeted molecule. Mol Biol Cell 19(3):1104–1112. https://doi.org/10.1091/mbc.E07-10-1004
Voellmy R, Boellmann F (2007) Chaperone regulation of the heat shock protein response. Adv Exp Med Biol 594:89–99. https://doi.org/10.1007/978-0-387-39975-1_9
Zhang Y, Ma C, Delohery T, Nasipak B, Foat BC, Bounoutas A, Bussemaker HJ, Kim SK, Chalfie M (2002) Identification of genes expressed in C. elegans touch receptor neurons. Nature 418(6895):331–335. https://doi.org/10.1038/nature00891
Acknowledgments
The N2 wildtype and OG497 strains were provided by the CGC, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). The authors acknowledge Dr. Martin Chalfie (Columbia University) for his helpful suggestions and advice.
Funding
This work was funded by NIH grant AG052149 to S.D.W. and M.W. was supported by NIH grant GM122522 to Martin Chalfie.
Author information
Authors and Affiliations
Contributions
A.D., M. W., and S.D.W. designed the study. A.D. and M.W. performed the experiments and the data analyses. A.D., M. W., and S.D.W. contributed to figure design. A.D. and S.D.W. wrote the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Figure S1
CRISPR/Cas9-mediated transgenesis of the endogenous hsf-1 locus in C. elegans to include a C-terminal GFP tag. (A) Approximate schematic cartoon model of modified HSF-1 locus with accompanying genotyping primers located within or outside of sequence included in HRT targeted genomic locus. of insertion of Homologous Repair Template (HRT) into endogenous HSF-1 genomic locus. Whole worm lysate was used to genotype for the appropriate CRISPR/Cas9 mediated knock-in compared to N2 (wildtype) and CRISPR strain SDW015 (HSF-1::GFP). (PPTX 140 kb).
Figure S2
HSF-1::GFP nuclear stress bodies can form throughout the C. elegans germline. Digitally stitched brightfield and fluorescence images of the germline of L4 CRISPR HSF-1::GFP (SDW015) are shown. Magnified inserts in the loop, distal, and proximal regions of the germline contain germ cells with the presence of HSF-1::GFP nSBs in the absence of any exogenous stressors. Yellow arrows indicate HSF-1::GFP nSBs. Scale bar in A-C represents 45 microns. Scale bar in D-F represents 5 microns. (G) Quantification of n = 10 L4 germlines for the presence of HSF-1::GFP nSBs. (PPTX 1117 kb).
Figure S3
–HSF-1::GFP forms nuclear stress bodies upon heat shock after exposure to acrylamide. (A-D) Confocal brightfield images of SDW015 shows expression of HSF-1::GFP during control conditions or upon exposure to acrylamide (7 mM/5 h) with or without heat shock for 5 min at 33 °C. (HS). (E) Nuclear stress body formation was quantified and graphed with n > 8 (n = number of animals assessed). All conditions were compared to Control –HS. (F) CL2166 (pgst-4:GFP) animals before and after a 5 hour exposure to 7 mM acrylamide shows that acrylamide induces the expected oxidative stress response. Scale bar in A-D presents 5 microns, scale bar in F represents 1000 microns. (PPTX 2122 kb).
Figure S4
Increased numbers of HSF-1::GFP nSBs are displayed throughout aging. (A) SDW015 animals were grown without heat shock and assessed for the number of nSBs within individual hypodermal nuclei beginning at the last larval stage (L4), in young adult (YA), fully gravid adults (GA) and then for the indicated number of days (+n) post GA. The number of nSBs per cell assessed was plotted in (A) with the mean number of nSBs per cell indicated (B). The red bar in (A) represents the mean number of nSBs present per cell. Approximately 300 individual cells were assessed across n = 8 individual animals per condition. Significance was determined with a One-way ANOVA followed by a Tukey Post-Hoc test of all comparisons. *** indicates p < 0.0001. All conditions were compared to L4, and GA was also compared to GA + 9 as indicated with brackets. (PPTX 125 kb).
Figure S5
PLM neurons form HSF-1 nuclear stress bodies in response to heat shock. Confocal fluorescence images of HSF-1::GFP CRISPR; pmec-17::RFP worms (SDW077) are shown without heat shock (A-C) and with heat shock (D-F). PLM neurons are marked by RFP. (G) The fraction of PLM nuclei scored as negative for nSBs or positive for nSBs for n ≥ 8 worms with or without heat shock was quantified and graphed for images (A-F). Significance indicated compares –HS to +HS, *** indicates p < 0.0001. Scale bar represents 5 microns. (PPTX 631 kb).
Figure S6
Adult nerve ring neurons do not form HSF-1 nuclear stress bodies upon the transition to adulthood. Confocal fluorescence images of gravid adult HSF-1::GFP CRISPR; pmec-17::RFP worms (SDW077) are shown. Nerve ring neurons are identified by the pharyngeal bulbs in differential interference contrast (DIC) microscopy and the ALM branch with the red TRN marker. Yellow dotted "nerve ring" box placement is around the location of the terminal pharyngeal bulb. Two examples are shown. Scale bar represents 10 microns. (PPTX 2359 kb).
Rights and permissions
About this article
Cite this article
Deonarine, A., Walker, M.W.G. & Westerheide, S.D. HSF-1 displays nuclear stress body formation in multiple tissues in Caenorhabditis elegans upon stress and following the transition to adulthood. Cell Stress and Chaperones 26, 417–431 (2021). https://doi.org/10.1007/s12192-020-01188-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12192-020-01188-9