A complex containing the CPSF73 endonuclease and other polyadenylation factors associates with U7 snRNP and is recruited to histone pre-mRNA for 3'-end processing

Xiao-Cui Yang; Ivan Sabath; Jan Dębski; Magdalena Kaus-Drobek; Michał Dadlez; William F Marzluff; Zbigniew Dominski

doi:10.1128/MCB.00653-12

A complex containing the CPSF73 endonuclease and other polyadenylation factors associates with U7 snRNP and is recruited to histone pre-mRNA for 3'-end processing

Mol Cell Biol. 2013 Jan;33(1):28-37. doi: 10.1128/MCB.00653-12. Epub 2012 Oct 15.

Authors

Xiao-Cui Yang¹, Ivan Sabath, Jan Dębski, Magdalena Kaus-Drobek, Michał Dadlez, William F Marzluff, Zbigniew Dominski

Affiliation

¹ Department of Biochemistry and Biophysics, Program in Molecular Biology and Biotechnology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

Abstract

Animal replication-dependent histone pre-mRNAs are processed at the 3' end by endonucleolytic cleavage that is not followed by polyadenylation. The cleavage reaction is catalyzed by CPSF73 and depends on the U7 snRNP and its integral component, Lsm11. A critical role is also played by the 220-kDa protein FLASH, which interacts with Lsm11. Here we demonstrate that the N-terminal regions of these two proteins form a platform that tightly interacts with a unique combination of polyadenylation factors: symplekin, CstF64, and all CPSF subunits, including the endonuclease CPSF73. The interaction is inhibited by alterations in each component of the FLASH/Lsm11 complex, including point mutations in FLASH that are detrimental for processing. The same polyadenylation factors are associated with the endogenous U7 snRNP and are recruited in a U7-dependent manner to histone pre-mRNA. Collectively, our studies identify the molecular mechanism that recruits the CPSF73 endonuclease to histone pre-mRNAs, reveal an unexpected complexity of the U7 snRNP, and suggest that in animal cells polyadenylation factors assemble into two alternative complexes-one specifically crafted to generate polyadenylated mRNAs and the other to generate nonpolyadenylated histone mRNAs that end with the stem-loop.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs
Animals
Apoptosis Regulatory Proteins / genetics
Apoptosis Regulatory Proteins / metabolism
Base Sequence
Calcium-Binding Proteins / genetics
Calcium-Binding Proteins / metabolism
Cleavage And Polyadenylation Specificity Factor / genetics
Cleavage And Polyadenylation Specificity Factor / metabolism*
Cleavage Stimulation Factor
HeLa Cells
Histones / metabolism*
Humans
Mice
Molecular Sequence Data
Mutation
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
RNA 3' End Processing*
RNA Precursors / metabolism*
RNA-Binding Proteins / genetics
RNA-Binding Proteins / metabolism
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Ribonucleoprotein, U7 Small Nuclear / genetics
Ribonucleoprotein, U7 Small Nuclear / metabolism*
mRNA Cleavage and Polyadenylation Factors / genetics
mRNA Cleavage and Polyadenylation Factors / metabolism*

Substances

Apoptosis Regulatory Proteins
CASP8AP2 protein, human
CSTF2T protein, human
Calcium-Binding Proteins
Cleavage And Polyadenylation Specificity Factor
Cleavage Stimulation Factor
Histones
Lsm11 protein, human
Nuclear Proteins
RNA Precursors
RNA-Binding Proteins
Recombinant Proteins
Ribonucleoprotein, U7 Small Nuclear
SYMPK protein, human
mRNA Cleavage and Polyadenylation Factors

Abstract

Publication types

MeSH terms

Substances

Grants and funding