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PMID: 1991453 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

A novel splicing factor is an integral component of 200S large nuclear ribonucleoprotein (InRNP) particles.

The EMBO journal ·Vol. 10 ·No. 2 ·1991-02-00 ·Pages 425-32

Ast G, Goldblatt D, Offen D, Sperling J, Sperling R

Abstract

In previous studies we have shown that nuclear transcripts of several pre-mRNAs can be released from nuclei of mammalian cells in the form of large nuclear ribonucleoprotein (InRNP) particles. By electron microscopy, these particles appeared as compact composite structures, 50 nm in diameter, which invariably sedimented at the 200S region in sucrose gradients. In order to identify putative protein splicing factors associated with the 200S InRNP particles, a panel of monoclonal antibodies directed against these particles were screened for their ability to inhibit splicing of pre-mRNA in vitro. In this study we have focused on a nuclear protein of 88 kd in molecular weight, which is an integral component of the InRNP complex and is recognized by monoclonal antibodies from a specific clone. This protein has been identified here as a novel splicing factor by, (i) antibody inhibition of splicing in vitro and (ii) depletion of splicing activity from HeLa cell nuclear extract after removing the 88 kd polypeptide by immunoadsorption, and complementation of the depleted activity with an affinity-purified 88 kd antigen. This splicing factor has further been shown to be required for the assembly of an active splicing complex.

MeSH Terms
Animals Antibodies, Monoclonal Cell Nucleus/metabolism,ultrastructure HeLa Cells/metabolism Humans Mice Mice, Inbred BALB C Mice, Inbred Strains/immunology Microscopy, Electron Molecular Weight RNA Precursors/genetics RNA Splicing Ribonucleoproteins/immunology,metabolism,ultrastructure Transcription, Genetic
Chemicals
Antibodies, Monoclonal RNA Precursors Ribonucleoproteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ast G
Department of Organic Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
Goldblatt D
Offen D
Sperling J
Sperling R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1991-02-00
Pages
425-32
Language
English
Region
England
NLM ID
8208664
PMCID
PMC452663
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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