TY - JOUR

T1 - The human ribosome can interact with the abasic site in mRNA via a specific peptide of the uS3 protein located near the mRNA entry channel

AU - Ochkasova, Anastasia S.

AU - Meschaninova, Maria I.

AU - Venyaminova, Aliya G.

AU - Ivanov, Anton V.

AU - Graifer, Dmitri M.

AU - Karpova, Galina G.

N1 - Copyright © 2018 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

PY - 2019/3/1

Y1 - 2019/3/1

N2 - The small subunit ribosomal protein uS3 is a critically important player in the ribosome-mRNA interactions during translation and has numerous functions not directly related to protein synthesis in eukaryotes. A peculiar feature of the human uS3 protein is the ability of its fragment 55–64 exposed on the 40S subunit surface near the mRNA entry channel to form cross-links with 3′-terminal dialdehyde derivatives of various unstructured RNAs and with abasic sites in single-stranded DNAs. Here we showed that the ability of the above uS3 fragment to cross-link to abasic sites in DNAs is inherent only in mature cytoplasmic 40S subunits, but not nuclear pre-40S particles, which implies that it may be relevant to the ribosome-mRNA interplay. To clarify this issue, we investigated interactions of human ribosomes with synthetic mRNA analogues bearing an abasic site protected by a photocleavable group at the 3′-termini. We found that these mRNA analogues can form specific complexes with 80S ribosomes and 40S subunits, where the undamaged upstream part of the analogue is fixed in the mRNA binding channel by interaction with the P-site tRNA, and the downstream part located outside the ribosome is cross-linked to the uS3 fragment 55–64. The yield of cross-links of the mRNA analogues was rather high when their undamaged parts were bound to the mRNA channel prior to deprotection of the abasic site enabling its covalent attachment to the 40S subunit via the uS3 protein, but not vice versa. Based on our findings, one can assume that abasic sites, which can occur in mRNAs due to oxidative stress and ageing, are able to interact directly with the uS3 fragment exposed on the 40S subunit surface near the mRNA entry channel during translation. Consequently, the 40S subunit can be considered as a potential mRNA quality controller.

AB - The small subunit ribosomal protein uS3 is a critically important player in the ribosome-mRNA interactions during translation and has numerous functions not directly related to protein synthesis in eukaryotes. A peculiar feature of the human uS3 protein is the ability of its fragment 55–64 exposed on the 40S subunit surface near the mRNA entry channel to form cross-links with 3′-terminal dialdehyde derivatives of various unstructured RNAs and with abasic sites in single-stranded DNAs. Here we showed that the ability of the above uS3 fragment to cross-link to abasic sites in DNAs is inherent only in mature cytoplasmic 40S subunits, but not nuclear pre-40S particles, which implies that it may be relevant to the ribosome-mRNA interplay. To clarify this issue, we investigated interactions of human ribosomes with synthetic mRNA analogues bearing an abasic site protected by a photocleavable group at the 3′-termini. We found that these mRNA analogues can form specific complexes with 80S ribosomes and 40S subunits, where the undamaged upstream part of the analogue is fixed in the mRNA binding channel by interaction with the P-site tRNA, and the downstream part located outside the ribosome is cross-linked to the uS3 fragment 55–64. The yield of cross-links of the mRNA analogues was rather high when their undamaged parts were bound to the mRNA channel prior to deprotection of the abasic site enabling its covalent attachment to the 40S subunit via the uS3 protein, but not vice versa. Based on our findings, one can assume that abasic sites, which can occur in mRNAs due to oxidative stress and ageing, are able to interact directly with the uS3 fragment exposed on the 40S subunit surface near the mRNA entry channel during translation. Consequently, the 40S subunit can be considered as a potential mRNA quality controller.

KW - Abasic (AP) site

KW - Cross-linking

KW - Human ribosome

KW - mRNA surveillance

KW - Ribosomal protein uS3

KW - Ribosome-associated mRNA quality control

KW - Synthetic mRNA analogues

KW - S3

KW - ELEMENTS

KW - DIRECT TRANSLATION

KW - SINGLE-STRANDED-DNA

KW - CYCLE

KW - Humans

KW - Peptides/chemistry

KW - RNA, Messenger/chemistry

KW - Ribosomal Proteins/chemistry

KW - Female

KW - Ribosome Subunits, Small, Eukaryotic/chemistry

UR - http://www.scopus.com/inward/record.url?scp=85059534527&partnerID=8YFLogxK

U2 - 10.1016/j.biochi.2018.12.015

DO - 10.1016/j.biochi.2018.12.015

M3 - Article

C2 - 30594661

AN - SCOPUS:85059534527

VL - 158

SP - 117

EP - 125

JO - Biochimie

JF - Biochimie

SN - 0300-9084

ER -