N-terminal region of Saccharomyces cerevisiae eRF3 is essential for the functioning of the eRF1/eRF3 complex beyond translation termination.
BACKGROUND
Termination of translation in eukaryotes requires two release factors, eRF1, which recognizes all three nonsense codons and facilitates release of the nascent polypeptide chain, and eRF3 stimulating translation termination in a GTP-depended manner. eRF3 from different organisms possess a highly conservative C region (eRF3C), which is responsible for the function in translation termination, and almost always contain the N-terminal extension, which is inessential and vary both in structure and length. In the yeast Saccharomyces cerevisiae the N-terminal region of eRF3 is responsible for conversion of this protein into the aggregated and functionally inactive prion form.


RESULTS
Here, we examined functional importance of the N-terminal region of a non-prion form of yeast eRF3. The screen for mutations which are lethal in combination with the SUP35-C allele encoding eRF3C revealed the sup45 mutations which alter the N-terminal domain of eRF1 and increase nonsense codon readthrough. However, further analysis showed that synthetic lethality was not caused by the increased levels of nonsense codon readthrough. Dominant mutations in SUP35-C were obtained and characterized, which remove its synthetic lethality with the identified sup45 mutations, thus indicating that synthetic lethality was not due to a disruption of interaction with proteins that bind to this eRF3 region.


CONCLUSIONS
These and other data demonstrate that the N-terminal region of eRF3 is involved both in modulation of the efficiency of translation termination and functioning of the eRF1/eRF3 complex outside of translation termination.
Urakov
Valery N
VN
Institute of Experimental Cardiology, Cardiology Research Center, Moscow, 121552, Russia. urakov@cardio.ru
Valouev
Igor A
IA
Kochneva-Pervukhova
Natalia V
NV
Packeiser
Anna N
AN
Vishnevsky
Alexander Yu
AY
Glebov
Oleg O
OO
Smirnov
Vladimir N
VN
Ter-Avanesyan
Michael D
MD
D000483:Alleles; D018388:Codon, Terminator / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010443:Peptide Chain Termination, Translational / Q000235:genetics* / Q000502:physiology; D010454:Peptide Termination Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011328:Prions / Q000235:genetics; D011485:Protein Binding / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics
D018388:Codon, Terminator; D010454:Peptide Termination Factors; D011328:Prions; C068068:SUP35 protein, S cerevisiae; C080775:SUP45 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C087971:peptide-chain-release factor 3