Evidence that a nuclear matrix protein participates in premessenger RNA splicing.
The role of nuclear matrix proteins in premessenger RNA splicing has been investigated using antibodies raised against isolated rat liver nuclear matrix and cross-reactive with a 65-kDa HeLa cell nuclear matrix protein (IGA-65). IGA-65 is an internal nuclear matrix component which can be solubilized as a component of nuclear splicing extracts, by the action of endogenous ribonucleases, EDTA, and DTT during extract preparation. Preincubation of splicing extract with antibodies against IGA-65 (anti-IGA-65) inhibited in vitro splicing of exogenous adenovirus precursor RNA. Furthermore, assembly of precursor RNA into active spliceosome complexes was inhibited by pretreatment of extracts with anti-IGA-65, suggesting a role for IGA-65 during early spliceosome assembly. The IGA-65 present in splicing extracts was distinguishable from known U-snRNP and hnRNP proteins on protein gels. Furthermore, electrophoresis of splicing extract on native gels indicated that IGA-65 was present in protein complexes different from those containing U-snRNPs or hnRNP C protein. The data support identification of complexes containing IGA-65 as nuclear factors involved in pre-mRNA splicing and, by extension, suggest a role for the nuclear matrix during processing in vivo.
Smith
H C
HC
Department of Pathology, University of Rochester, New York 14642.
Harris
S G
SG
Zillmann
M
M
Berget
S M
SM
D000906:Antibodies; D034961:Antigens, Nuclear; D001324:Autoantigens / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism*; D003429:Cross Reactions; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics*; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing
D000906:Antibodies; D034961:Antigens, Nuclear; D001324:Autoantigens; D009687:Nuclear Proteins; D012322:RNA Precursors