DOI: 10.1046/j.1432-1327.2001.02177.x. The polypeptide chain release factor eRF1 specifically contacts the s<sup>4</sup>UGA stop codon located in the A site of eukaryotic ribosomes

The polypeptide chain release factor eRF1 specifically contacts the s4UGA stop codon located in the A site of eukaryotic ribosomes

10.1046/j.1432-1327.2001.02177.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

It has been shown previously [Brown, C.M. & Tate, W.P. (1994) J. Biol. Chem.269, 33164–33170.] that the polypeptide chain release factor RF2 involved in translation termination in prokaryotes was able to photocrossreact with mini‐messenger RNAs containing stop signals in which U was replaced by 4‐thiouridine (s4U). Here, using the same strategy we have monitored photocrosslinking to eukaryotic ribosomal components of 14‐mer mRNA in the presence of , and 42‐mer mRNA in the presence of tRNAAsp (tRNAAsp gene transcript). We show that: (a) both 14‐mer and 42‐mer mRNAs crossreact with ribosomal RNA and ribosomal proteins. The patterns of the crosslinked ribosomal proteins are similar with both mRNAs and sensitive to ionic conditions; (b) the crosslinking patterns obtained with 42‐mer mRNAs show characteristic modification upon addition of tRNAAsp providing evidence for appropriate mRNA phasing onto the ribosome. Similar changes are not detected with the 14‐mer pairs; (c) when eukaryotic polypeptide chain release factor 1 (eRF1) is added to the ribosome·tRNAAsp complex it crossreacts with the 42‐mer mRNA containing the s4UGA stop codon located in the A site, but not with the s4UCA sense codon; this crosslink involves the N‐terminal and middle domains of eRF1 but not the C domain which interacts with eukaryotic polypeptide chain release factor 3 (eRF3); (d) addition of eRF3 has no effect on the yield of eRF1–42‐mer mRNA crosslinking and eRF3 does not crossreact with 42‐mer mRNA. These experiments delineate the in vitro conditions allowing optimal phasing of mRNA on the eukaryotic ribosome and demonstrate a direct and specific contact of ‘core’ eRF1 and s4UGA stop codon within the ribosomal A site.

Bibliography

Chavatte, L., Frolova, L., Kisselev, L., & Favre, A. (2001). The polypeptide chain release factor eRF1 specifically contacts the s4UGA stop codon located in the A site of eukaryotic ribosomes. European Journal of Biochemistry, 268(10), 2896â2904. Portico.

Authors 4

Laurent Chavatte (first)
Ludmila Frolova (additional)
Lev Kisselev (additional)
Alain Favre (additional)

References 41 Referenced 49

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Dates

Type	When
Created	22 years, 5 months ago (March 11, 2003, 1:09 p.m.)
Deposited	1 year, 9 months ago (Oct. 31, 2023, 12:31 a.m.)
Indexed	1 month, 1 week ago (July 11, 2025, 6:29 a.m.)
Issued	24 years, 3 months ago (May 15, 2001)
Published	24 years, 3 months ago (May 15, 2001)
Published Online	23 years, 8 months ago (Dec. 20, 2001)
Published Print	24 years, 3 months ago (May 15, 2001)

Funders 0

None

BibTeX

@article{Chavatte_2001, title={The polypeptide chain release factor eRF1 specifically contacts the s4UGA stop codon located in the A site of eukaryotic ribosomes}, volume={268}, ISSN={1432-1033}, url={http://dx.doi.org/10.1046/j.1432-1327.2001.02177.x}, DOI={10.1046/j.1432-1327.2001.02177.x}, number={10}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Chavatte, Laurent and Frolova, Ludmila and Kisselev, Lev and Favre, Alain}, year={2001}, month=may, pages={2896–2904} }

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