MUTATIONS IN C-PART OF SUP35 INFLUENCE PROPERTIES OF [PSI+] FACTOR IN YEASTS

O.M. Zemlyanko, E.M. Maksutenko, N.P. Trubitcina, T.M. Rogoza, E.I. Porfirieva, G.A. Zhouravleva

Research output

3 Downloads (Pure)

Abstract

The cytoplasmic [PSI+] factor is one of the best characterized yeast prions. [PSI+] is formed by Sup35 protein aggregates. Sup35p is encoded by the SUP35 gene. It consists of the nonessential N-terminal [PSI+] prion forming domain; the charged middle (M) domain, which maintains [PSI+], and the C-terminal domain with GTP-binding sites essential for translation termination activity. Mutations in the SUP35 gene leads to nonsense suppression, [PSI+] have the same effect. Previously it was shown that mutations in N-terminal domain of Sup35p affect [PSI+] appearance and maintenance. However, it has been demonstrated that mutations in Sup35p C-terminal domain can also change [PSI+] properties. In this work, we have studied sup35-228, sup35-10, and sup35-25 mutations located in the GTP-binding region of Sup35p. Using SDD-AGE, we have shown that [PSI+] is lost in the presence of only mutant alleles (sup35-228, sup35-10, sup35-25). Using fluorescent microscopy we additionally demonstrated that sup35-228 do not maintain pre-existing [PSI+]. However, several clones were obtained (less than 10%), which retained the [PSI+] factor, but at the same time replaced the mutant sup35 allele with the wild-type allele. Our results indicate that [PSI+] is incompatible with mutations in the C-terminal domain of Sup35p, but reasons of this effect remained unclear. We supposed that this phenomenon may be caused by the [PSI+] destabilization or inability of mutant Sup35p to form and maintain prion aggregates. In order to test this hypothesis we analyzed properties of missense-mutant proteins in vitro. We have shown that mutated Sup35 proteins form aggregates with high molecular weight and their protein aggregation rate is faster relative to the wild-type Sup35p. However, the amyloid nature of these aggregates requires confirmation. Another reason for incompatibility of sup35 missense-mutations and [PSI+] may be significant translation termination impairment. [PSI+] sequestrate functional Sup35p into prion aggregates, this possibly leads to dramatic reduction in accuracy of translation termination and cell death. Taken together, our data suggest that the non-prion C-domain of Sup35p is involved in the process of [PSI +] appearance and maintenance. This work was supported by grant from RSF 18-14-00050, RFBR 17-54-150002, technical help was provided by Resource Center «Development of Molecular and Cell Technologies»
Original languageEnglish
Title of host publicationVII Съезд Вавиловского общества генетиков и селекционеров (ВОГиС)
Subtitle of host publicationСборник тезисов
Place of PublicationСПб.
PublisherИздательство «ВВМ»
Pages650
ISBN (Print)9785965112371
Publication statusPublished - 2019
EventМеждународный конгресс «VII съезд Вавиловского общества генетиков и селекционеров, посвященный 100-летию кафедры генетики СПбГУ, и ассоциированные симпозиумы»: ВОГиС 2019 - Санкт-Петербург
Duration: 18 Jun 201922 Jun 2019
https://events.spbu.ru/events/genetic-selection-2019

Conference

ConferenceМеждународный конгресс «VII съезд Вавиловского общества генетиков и селекционеров, посвященный 100-летию кафедры генетики СПбГУ, и ассоциированные симпозиумы»
Abbreviated titleВОГиС 2019
CountryRussian Federation
CityСанкт-Петербург
Period18/06/1922/06/19
Internet address

Cite this