PXD031227

PXD031227 is an original dataset announced via ProteomeXchange.

Title	Reconstitution of 3’-processing of mammalian pre-mRNA reveals a central role of RBBP6
Description	The 3’ ends of almost all eukaryotic mRNAs are generated in an essential two-step processing reaction, endonucleolytic cleavage of an extended precursor followed by the addition of a poly(A) tail. By reconstituting the reaction from overproduced and purified proteins, we provide a minimal list of fourteen polypeptides essential and two stimulatory for RNA processing. In a reaction depending on the polyadenylation signal AAUAAA, the reconstituted system cleaves pre-mRNA at a single preferred site corresponding to the one used in vivo. Among the proteins, Cleavage Factor I stimulates cleavage but is not essential, consistent with its prominent role in alternative polyadenylation. RBBP6 is required, structural data showing it to contact and presumably activate the endonuclease CPSF73 through its DWNN domain. The C-terminal domain of RNA polymerase II is dispensable. ATP, but not its hydrolysis, supports RNA cleavage by binding to the hClp1 subunit of cleavage factor II with submicromolar affinity.
HostingRepository	PRIDE
AnnounceDate	2022-03-04
AnnouncementXML	Submission_2022-03-04_04:52:22.746.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Florian Kluge
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-25 00:54:58	ID requested
⏵ 1	2022-03-04 04:52:23	announced

Schmidt M, Kluge F, Sandmeir F, K, ü, hn U, Sch, ä, fer P, T, ü, ting C, Ihling C, Conti E, Wahle E, Reconstitution of 3' end processing of mammalian pre-mRNA reveals a central role of RBBP6. Genes Dev, 36(3-4):195-209(2022) [pubmed]

submitter keyword: XL-MS , 3’-processing , RBBP6 , CPSF

Elmar Wahle
contact affiliation	University of Halle Department of Biochemistry Germany
contact email	ewahle@biochemtech.uni-halle.de
lab head
Florian Kluge
contact affiliation	Institute for Biochemistry and Biotechnology; AG Wahle
contact email	florian.kluge@bct.uni-halle.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/03/PXD031227

PRIDE project URI

• PRIDE
  1. PXD031227
     1. Label: PRIDE project
     2. Name: Reconstitution of 3’-processing of mammalian pre-mRNA reveals a central role of RBBP6