Proteomics

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Spliceosome specifically associates with CTD phospho-serine 5 isoform of RNA polymerase II to promote co-transcriptional splicing


ABSTRACT: Co-transcriptional splicing of introns is a defining feature of eukaryotic gene expression. We show that the mammalian spliceosome specifically associates with the S5P CTD isoform of RNA polymerase II (Pol II) as it elongates across spliced exons of protein coding genes, both in human Hela and murine lymphoid cell lines. Immuno-precipitation of MNase digested chromatin with phospho CTD specific antibodies reveals that components of the active spliceosome (both snRNA and proteins) form a specific complex with S5P CTD Pol II. Furthermore a dominant splicing intermediate formed by cleavage at intron 5’ss results in the tethering of upstream exons to this complex at all spliced exons. These are invariably connected to upstream spliced constitutive and less frequently to alternative exons. Finally S5P CTD Pol II accumulates over spliced exons but not adjacent introns. We propose that mammalian splicing employs a rapid, co-transcriptional splicing mechanism based on CTD phosphorylation transitions.

INSTRUMENT(S): LTQ XL

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell Of Cervix, Hela Cell

DISEASE(S): Cervical Cancer

SUBMITTER: Kenny Rebelo  

LAB HEAD: Maria Carmo Fonseca

PROVIDER: PXD008197 | Pride | 2018-10-05

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
CPFP_MSS10615_19062015.mzid Mzid
CPFP_MSS10622_22062015.mzid Mzid
CPFP_MSS10636_01072015.mzid Mzid
Qex03_BT_150619_Taka_Ch1.mgf Mgf
Qex03_BT_150619_Taka_Ch1.raw Raw
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