Interaction of the human autoantigen p150 with splicing snRNPs
10.1242/jcs.105.3.685
Crossref journal-article
The Company of Biologists
Journal of Cell Science (237)
Abstract

ABSTRACT An important goal of studies on pre-mRNA splicing is to identify factors that mediate the snRNP-snRNP and snRNP-pre-mRNA interactions that take place in the spliceosome. The U4/U6 snRNP is one of the four snRNPs that are subunits of spliceosomes. A rare patient autoimmune serum (MaS serum) has recently been identified that specifically immunoprecipitates U4/U6 snRNP from HeLa cell extracts through recognition of a 150 kDa autoantigen (p150) (Okano and Medsger, Journal of Immunology, 146, 535-542, 1991). Here we show that in addition to U4/U6 snRNP, p150 can also be detected associated with 20 S U5, U4/U6.U5 and 17 S U2 snRNPs, but not with U1 snRNP. In each particle p150 is present in sub-stoichiometric levels relative to the major snRNP proteins. We show that MaS serum selectively immunoprecipitates a sub-population of U4/U6 snRNPs in which the m3G-cap structure is masked and that p150 is preferentially associated with U6 snRNA in the U4/U6 particle. Anti-p150 antibodies show widespread nucleoplasmic staining, excluding nucleoli, with an elevated concentration in coiled bodies. This changes to a discrete punctate pattern when cells are treated with α-amanitin. Both the cytological and biochemical data indicate that the p150 autoantigen is a snRNP-associated factor in vivo. We also present biochemical evidence confirming that assembly of U4/U6 and U5 snRNPs into a U4/U6.U5 tri-snRNP particle is an integral step in the spliceosome assembly pathway. Addition of the purified U4/U6.U5 tri-snRNP restores splicing activity to inactivated HeLa nuclear extracts in which splicing had been inhibited by specific depletion of either the U4/U6 or U5 snRNPs.

Bibliography

Blencowe, B. J., Carmo-Fonseca, M., Behrens, S.-E., Lührmann, R., & Lamond, A. I. (1993). Interaction of the human autoantigen p150 with splicing snRNPs. Journal of Cell Science, 105(3), 685–697.

Authors 5
  1. Benjamin J. Blencowe (first)
  2. Maria Carmo-Fonseca (additional)
  3. Sven-Erik Behrens (additional)
  4. Reinhard Lührmann (additional)
  5. Angus I. Lamond (additional)
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Dates
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Created 4 years, 4 months ago (April 25, 2021, 11:34 a.m.)
Deposited 1 year, 8 months ago (Dec. 23, 2023, 4:57 p.m.)
Indexed 4 months, 2 weeks ago (April 8, 2025, 10:21 p.m.)
Issued 32 years, 1 month ago (July 1, 1993)
Published 32 years, 1 month ago (July 1, 1993)
Published Online 32 years, 1 month ago (July 1, 1993)
Published Print 32 years, 1 month ago (July 1, 1993)
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@article{Blencowe_1993, title={Interaction of the human autoantigen p150 with splicing snRNPs}, volume={105}, ISSN={1477-9137}, url={http://dx.doi.org/10.1242/jcs.105.3.685}, DOI={10.1242/jcs.105.3.685}, number={3}, journal={Journal of Cell Science}, publisher={The Company of Biologists}, author={Blencowe, Benjamin J. and Carmo-Fonseca, Maria and Behrens, Sven-Erik and Lührmann, Reinhard and Lamond, Angus I.}, year={1993}, month=jul, pages={685–697} }