Recognition but no repair of abasic site in single-stranded DNA by human ribosomal uS3 protein residing within intact 40S subunit.
Isolated human ribosomal protein uS3 has extra-ribosomal functions including those related to base excision DNA repair, e.g. AP lyase activity that nicks double-stranded (ds) DNA 3΄ to the abasic (AP) site. However, the ability of uS3 residing within ribosome to recognize and cleave damaged DNA has never been addressed. Here, we compare interactions of single-stranded (ss) DNA and dsDNA bearing AP site with human ribosome-bound uS3 and with the isolated protein, whose interactions with ssDNA were not yet studied. The AP lyase activity of free uS3 was much higher with ssDNA than with dsDNA, whereas ribosome-bound uS3 was completely deprived of this activity. Nevertheless, an exposed peptide of ribosome-bound uS3 located far away from the putative catalytic center previously suggested for isolated uS3 cross-linked to full-length uncleaved ssDNA, but not to dsDNA. In contrast, free uS3 cross-linked mainly to the 5΄-part of the damaged DNA strand after its cleavage at the AP site. ChIP-seq analysis showed preferential uS3 binding to nucleolus-associated chromatin domains. We conclude that free and ribosome-bound uS3 proteins interact with AP sites differently, exhibiting their non-translational functions in DNA repair in and around the nucleolus and in regulation of DNA damage response in looped DNA structures, respectively.
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.
Anastasia S
AS
Grosheva
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.|Novosibirsk State University, Novosibirsk 630090, Russia.
Dmitry O
DO
Zharkov
Max-Delbrück-Center for Molecular Medicine, D-13092 Berlin, Germany.
Joachim
J
Stahl
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.|Novosibirsk State University, Novosibirsk 630090, Russia.
Alexander V
AV
Gopanenko
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.
Alexey E
AE
Tupikin
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.
Marsel R
MR
Kabilov
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.|Novosibirsk State University, Novosibirsk 630090, Russia.
Dmitri M
DM
Graifer
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk 630090, Russia.|Novosibirsk State University, Novosibirsk 630090, Russia.
Galina G
GG
Karpova
D002503:Centromere; D002877:Chromosomes, Human / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D000072417:Protein Domains; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism*; D054682:Ribosome Subunits, Small, Eukaryotic / Q000378:metabolism*
D004277:DNA, Single-Stranded; C578290:RPS3 protein, human; D012269:Ribosomal Proteins