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XL-MS of RBBP6-bound CPSF complex

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Felix Sandmeir

LAB HEAD: Elena Conti

PROVIDER: PXD031242 | Pride | 2022-03-07

REPOSITORIES: Pride

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			Action	DRS
	20210716_coreQEP1_NiKr_LC12-18_SA_Orbi3928_Sample03_Fraktion01.raw	Raw
	20210716_coreQEP1_NiKr_LC12-18_SA_Orbi3928_Sample03_Fraktion01_Crosslinks.txt	Txt
	20210716_coreQEP1_NiKr_LC12-18_SA_Orbi3928_Sample03_Fraktion02.raw	Raw
	20210716_coreQEP1_NiKr_LC12-18_SA_Orbi3928_Sample03_Fraktion03.raw	Raw
	20210716_coreQEP1_NiKr_LC12-18_SA_Orbi3928_Sample03_Fraktion04.raw	Raw

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