TY - JOUR

T1 - Human ribosomal protein eS1 is engaged in cellular events related to processing and functioning of U11 snRNA

AU - Gopanenko, Alexander V.

AU - Malygin, Alexey A.

AU - Tupikin, Alexey E.

AU - Laktionov, Pavel P.

AU - Kabilov, Marsel R.

AU - Karpova, Galina G.

N1 - © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2017/9/6

Y1 - 2017/9/6

N2 - Ribosomal proteins are involved in many cellular processes through interactions with various RNAs. Here, applying the photoactivatable-ribonucleosideenhanced cross-linking and immunoprecipitation approach to HEK293 cells overproducing ribosomal protein (rp) eS1, we determined the products of RNU5A-1 and RNU11 genes encoding U5 and U11 snRNAs as the RNA partners of ribosome-unbound rp eS1. U11 pre-snRNA-associated rp eS1 was revealed in the cytoplasm and nucleus where rp eS1- bound U11/U12 di-snRNP was also found. Utilizing recombinant rp eS1 and 4-thiouridine-containing U11 snRNA transcript, we identified an N-terminal peptide contacting the U-rich sequence in the Sm sitecontaining RNA region. We also showed that the rp eS1 binding site on U11 snRNA is located in the cleft between stem-loops I and III and that its structure mimics the respective site on the 18S rRNA. It was found that cell depletion of rp eS1 leads to a decrease in the splicing efficiency of minor introns and to an increase in the level of U11 pre-snRNA with the unprocessed 3″ terminus. Our findings demonstrate the engagement of human rp eS1 in events related to the U11 snRNA processing and to minor-class splicing. Contacts of rp eS1 with U5 snRNA in the minor precatalytic spliceosome are discussed.

AB - Ribosomal proteins are involved in many cellular processes through interactions with various RNAs. Here, applying the photoactivatable-ribonucleosideenhanced cross-linking and immunoprecipitation approach to HEK293 cells overproducing ribosomal protein (rp) eS1, we determined the products of RNU5A-1 and RNU11 genes encoding U5 and U11 snRNAs as the RNA partners of ribosome-unbound rp eS1. U11 pre-snRNA-associated rp eS1 was revealed in the cytoplasm and nucleus where rp eS1- bound U11/U12 di-snRNP was also found. Utilizing recombinant rp eS1 and 4-thiouridine-containing U11 snRNA transcript, we identified an N-terminal peptide contacting the U-rich sequence in the Sm sitecontaining RNA region. We also showed that the rp eS1 binding site on U11 snRNA is located in the cleft between stem-loops I and III and that its structure mimics the respective site on the 18S rRNA. It was found that cell depletion of rp eS1 leads to a decrease in the splicing efficiency of minor introns and to an increase in the level of U11 pre-snRNA with the unprocessed 3″ terminus. Our findings demonstrate the engagement of human rp eS1 in events related to the U11 snRNA processing and to minor-class splicing. Contacts of rp eS1 with U5 snRNA in the minor precatalytic spliceosome are discussed.

KW - Amino Acid Sequence

KW - Base Sequence

KW - Escherichia coli/genetics

KW - HEK293 Cells

KW - Humans

KW - Introns

KW - Models, Molecular

KW - Nucleic Acid Conformation

KW - Protein Biosynthesis

KW - Protein Conformation, alpha-Helical

KW - Protein Interaction Domains and Motifs

KW - RNA Splicing

KW - RNA, Ribosomal, 18S/genetics

KW - RNA, Small Nuclear/chemistry

KW - Ribonucleoproteins, Small Nuclear/genetics

KW - Ribosomal Proteins/chemistry

KW - Ribosomes/genetics

KW - Spliceosomes/genetics

KW - Thiouridine/metabolism

KW - ENTRY SITE

KW - PRE-MESSENGER-RNA

KW - INTERACTS

KW - 5 SPLICE-SITE

KW - 40S RIBOSOME

KW - 80S RIBOSOME

KW - MECHANISM

KW - BINDING

KW - EXPRESSION

KW - INSIGHTS

UR - http://www.scopus.com/inward/record.url?scp=85031048737&partnerID=8YFLogxK

U2 - 10.1093/nar/gkx559

DO - 10.1093/nar/gkx559

M3 - Article

C2 - 28666385

AN - SCOPUS:85031048737

VL - 45

SP - 9121

EP - 9137

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 15

ER -