The role of DAF-41 in lifespan of Caenorhabditis elegans

master's thesis

• Title (croatian): Uloga DAF-41 u životnom vijeku Caenorhabditis elegans
• Author: Margareta Sačer
• Mentor: Adam Antebi (mentor)
• Mentor: Dubravka Hranilović (komentor)
• Committee member: Dubravka Hranilović (predsjednik povjerenstva)
• Committee member: Biljana Balen (član povjerenstva)
• Committee member: Dunja Leljak-Levanić (član povjerenstva)
• Committee member: Inga Urlić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2024-07-15, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Tiku et al. (2017) published that loss of the protein NCL-1 (abnormal NuCLeoli 1) abolishes
the extended lifespan in major Caenorhabditis elegans longevity pathways, including the
germline-deficient glp-1 (abnormal Germ Line Proliferation 1) C. elegans mutant. Recent
unpublished work from the Antebi group identified a protein DAF-41 (Abnormal Dauer
Formation 41), an HSP-90 (heat shock protein 90) co-chaperone, as a potential downstream
regulator of glp-1ncl-1 lifespan. I investigated the role of daf-41 in glp-1ncl-1 lifespan, finding
that daf-41 downregulation extended the lifespan of glp-1ncl-1 worms. Moreover, I performed
RT-qPCR analysis of genes associated with the insulin signaling pathway (daf-16, lipl-4, sod3, dod-3), heat-shock response (hsf-1, hsp-70, hsp-16.2, hsp-4), and steroid pathway (daf-12,
fard-1, cdr-6, daf-36), quantified GFP-tagged HSF-1 and DAF-16 transcription factor activity
and measured thermotolerance and oxidative stress resistance in different mutant backgrounds.
Changes in gene expression, DAF-16 and HSF-1 activity, thermotolerance, and oxidative stress
resistance did not clearly explain the mechanism behind this lifespan extension.

Abstract (english)

Tiku i suradnici (2017) primijetili su da delecija gena tumor-supresora NCL-1 (eng. Nucleoli
1) skraćuje produljeni životni vijek u više dugovječnih modela Caenorhabditis elegans, među
kojima i u dvostrukom mutantu glp-1ncl-1 (eng. Abnormal Germ Line Proliferation 1).
Prethodno neobjavljena istraživanja Antebija i suradnika nedavno su identificirala protein
DAF-41 (eng. Abnormal Dauer Formation 41), ko-šaperon proteina HSP-90 (eng. Heat Shock
Protein 90) kao potencijalnog nizvodnog regulatora životnog vijeka mutanta glp-1ncl-1.
Otkrila sam da utišavanje gena daf-41 produžuje životni vijek C. elegans mutanta glp-1ncl-1.
Kako bih otkrila potencijalni signalni put ovog fenomena, provela sam RT-qPCR analizu gena
inzulinskog signalnog puta (daf-16, lipl-4, sod-3, dod-3), odgovora na toplinski šok (hsf-1, hsp70, hsp-16.2, hsp-4) i steroidnog signalnog puta (daf-12, fard-1, cdr-6, daf-36). Zatim sam
kvantificirala aktivnost fluorescentno obilježenih transkripcijskih faktora HSF-1 i DAF-16 te
mjerila otpornost oblića na toplinski i oksidativni stres. Međutim, promjene u ekspresiji gena,
aktivnosti DAF-16 i HSF-1, otpornost na toplinski i oksidativni stres nisu dali jasan uvid u
mehanizam produženja životnog vijeka mutanata glp-1ncl-1 s utišanim genom daf-41.

• Keywords: HSP-90; p23; GLP-1; NCL-1; DAF-16; HSF-1
• Keywords (english): HSP-90; p23; GLP-1; NCL-1; DAF-16; HSF-1
• Language: english
• URN:NBN: urn:nbn:hr:217:564872
• Study programme: Title: Molecular Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra molekularne biologije (sveučilišni/a magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-08-26 13:24:14