RT Journal Article
SR Electronic
T1 RBBP6 activates the pre-mRNA 3’-end processing machinery in humans
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.11.02.466915
DO 10.1101/2021.11.02.466915
A1 Vytaute Boreikaite
A1 Thomas Elliott
A1 Jason Chin
A1 Lori A Passmore
YR 2021
UL http://biorxiv.org/content/early/2021/11/02/2021.11.02.466915.abstract
AB 3’-end processing of most human mRNAs is carried out by the cleavage and polyadenylation specificity factor (CPSF; CPF in yeast). Endonucleolytic cleavage of the nascent pre-mRNA defines the 3’-end of the mature transcript, which is important for mRNA localization, translation and stability. Cleavage must therefore be tightly regulated. Here, we reconstitute specific and efficient 3’-endonuclease activity of human CPSF with purified proteins. This requires the sevensubunit CPSF as well as three additional protein factors: cleavage stimulatory factor (CStF), cleavage factor IIm (CFIIm) and, importantly, the multi-domain protein RBBP6. Unlike its yeast homologue Mpe1, which is a stable subunit of CPF, RBBP6 does not copurify with CPSF and is recruited in an RNA-dependent manner. Sequence and mutational analyses suggest that RBBP6 interacts with the WDR33 and CPSF73 subunits of CPSF. Thus, it is likely that the role of RBBP6 is conserved from yeast to human. Overall, our data are consistent with CPSF endonuclease activation and site-specific pre-mRNA cleavage being highly controlled to maintain fidelity in RNA processing.Competing Interest StatementThe authors have declared no competing interest.