DOI: 10.1073/pnas.94.2.385. Identification of the proteins of the yeast U1 small nuclear ribonucleoprotein complex by mass spectrometry

Identification of the proteins of the yeast U1 small nuclear ribonucleoprotein complex by mass spectrometry

10.1073/pnas.94.2.385

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Here we report the rapid identification of the proteins of the spliceosomal U1 small nuclear ribonucleoprotein (snRNP) from the yeast Saccharomyces cerevisiae by searching mass spectrometric data in genomic sequence databases. The U1 snRNP, containing a histidine-tagged 70K protein, was isolated from cell extracts by anti m 3 G-cap immunoaffinity and subsequent nickel nitrilotriacetic acid chromatography. A U1 snRNP fraction containing 20 proteins was obtained. Further purification by glycerol gradient centrifugation identified nine U1 snRNP specific and six common proteins. The U1 snRNP proteins were partially sequenced by nanoelectrospray mass spectrometry, and their genes were identified in the data base via multiple peptide sequence tags. Apart from the already known common proteins D1, D3, F, and G, the D2 and E homologs were also identified. The same six common proteins were detected in core U2 snRNP, which was purified and analyzed separately. The biochemical association of these six proteins with yeast snRNPs is shown here for the first time. Intriguingly, the Sm B/B′ homolog was not detected. In addition to the well characterized yeast U1 specific proteins [U1-70K (Snp1p), U1-A (Mud1p), Prp39p, and Prp40p] the homolog of the U1-C protein was identified together with four additional novel U1 specific proteins, which are not found in mammalian U1. This is the first time that the components of a multiprotein complex from an organism with a sequenced genome have been characterized by mass spectrometry. The technique should be applicable to any protein complex that can be biochemically purified from an organism whose genome is known.

Bibliography

Neubauer, G., Gottschalk, A., Fabrizio, P., SÃ©raphin, B., LÃ¼hrmann, R., & Mann, M. (1997). Identification of the proteins of the yeast U1 small nuclear ribonucleoprotein complex by massâspectrometry. Proceedings of the National Academy of Sciences, 94(2), 385â390.

Authors 6

Gitte Neubauer (first)
Alexander Gottschalk (additional)
Patrizia Fabrizio (additional)
Bertrand Séraphin (additional)
Reinhard Lührmann (additional)
Matthias Mann (additional)

References 26 Referenced 150

S D Patterson, R Aebersold Electrophoresis 16, 791–814 (1995). / Electrophoresis by Patterson S D (1995)
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10.1021/ac00096a002
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10.1038/379466a0
B C Rymond, M Rosbash The Molecular and Cellular Biology of the Yeast Saccaromyces, eds E W Jones, J R Pringle, J R Broach (Cold Spring Harbor Lab. Press, Plainview, NY) I, 143–192 (1992). / The Molecular and Cellular Biology of the Yeast Saccaromyces by Rymond B C (1992)
M J Moore, C C Query, P A Sharp The RNA World, eds R F Gesteland, J F Atkins (Cold Spring Harbor Lab. Press, Plainview, NY), pp. 303–357 (1993). / The RNA World by Moore M J (1993)
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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:31 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 3:18 p.m.)
Indexed	1 month ago (July 24, 2025, 7:52 a.m.)
Issued	28 years, 7 months ago (Jan. 21, 1997)
Published	28 years, 7 months ago (Jan. 21, 1997)
Published Online	28 years, 7 months ago (Jan. 21, 1997)
Published Print	28 years, 7 months ago (Jan. 21, 1997)

Funders 0

None

BibTeX

@article{Neubauer_1997, title={Identification of the proteins of the yeast U1 small nuclear ribonucleoprotein complex by mass spectrometry}, volume={94}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.94.2.385}, DOI={10.1073/pnas.94.2.385}, number={2}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Neubauer, Gitte and Gottschalk, Alexander and Fabrizio, Patrizia and Séraphin, Bertrand and Lührmann, Reinhard and Mann, Matthias}, year={1997}, month=jan, pages={385–390} }

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