PXD014164-1

PXD014164 is an original dataset announced via ProteomeXchange.

Title	Composition and processing activity of a semi-recombinant holo U7 snRNP
Description	In animal cells, replication-dependent histone pre-mRNAs are cleaved at the 3’ end by U7 snRNP consisting of two core components: a ~60-nucleotide U7 snRNA and a ring of seven proteins, with Lsm10 and Lsm11 replacing the spliceosomal SmD1 and SmD2. Lsm11 interacts with FLASH and together they recruit the endonuclease CPSF73 and other polyadenylation factors, forming catalytically active holo U7 snRNP. Here, we assembled core U7 snRNP bound to FLASH from recombinant components and analyzed its appearance by electron microscopy and ability to support histone pre-mRNA processing in the presence of polyadenylation factors from nuclear extracts. We demonstrate that semi-recombinant holo U7 snRNP reconstituted in this manner has the same composition and functional properties as endogenous U7 snRNP, being capable of accurately cleaving histone pre-mRNAs in a reconstituted in vitro processing reaction. We also demonstrate that the U7-specific Sm ring assembles efficiently in vitro on a spliceosomal Sm site but the resultant semi-recombinant holo U7 snRNP fails to accurately cleave histone pre-mRNAs. This approach offers a unique opportunity to create engineered U7 snRNPs to study the role of various regions in the Sm proteins and U7 snRNA in the biogenesis of a functional holo U7 snRNP.
HostingRepository	PRIDE
AnnounceDate	2019-11-25
AnnouncementXML	Submission_2019-11-25_00:54:59.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Katarzyna Bucholc
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-06-06 09:04:22	ID requested
⏵ 1	2019-11-25 00:55:00	announced
2	2019-12-16 01:01:37	announced	2019-12-16: Updated publication reference for PubMed record(s): 31819999.
3	2024-10-22 04:55:30	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: U7 snRNP, Sm proteins, Lsm11, FLASH, histone pre-mRNA, 3’ end processing, CPSF73

Michał Dadlez
contact affiliation	Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics PAS
contact email	michald@ibb.waw.pl
lab head
Katarzyna Bucholc
contact affiliation	Institute of Biochemistry and Biophysics, Warsaw
contact email	kabucholc@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD014164
     1. Label: PRIDE project
     2. Name: Composition and processing activity of a semi-recombinant holo U7 snRNP