Structure-probing of U1 SnRNPs Gradually Depleted of the U1-specific Proteins A, C and 70k. Evidence That A Interacts Differentially with Developmentally Regulated Mouse U1 SnRNA Variants

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1990 Feb 11

PMID 2137909

Citations 31

Authors

M Bach

A Krol

R Luhrmann

Affiliations

Soon will be listed here.

Abstract

The interaction of the U1-specific proteins 70k, A and C with U1 snRNP was studied by depleting gradually U1 snRNPs of the U1-specific proteins by Mono-Q chromatography at elevated temperatures (20-37 degrees C). U1 snRNP species were obtained which were selectively depleted of either protein C, A, C and A, or of all three U1-specific proteins C, A and 70k while retaining the common proteins B' to G. These various types of U1 snRNP particles were used to study the differential accessibility of defined regions of U1 RNA towards nucleases V1 and S1 dependent on the U1 snRNP protein composition. The data indicate that in the U1 snRNP protein 70k interacts with stem/loop A and protein A with stem/loop B of U1 RNA. The presence or absence of protein C did not affect the nuclease digestion patterns of U1 RNA. Our results suggest further that the binding of protein A to the U1 snRNP particle should be independent of proteins 70k and C. Mouse cells contain two U1 RNA species, U1a and U1b, which differ in the structure of stem/loop B, with U1a exhibiting the same stem/loop B sequence as U1 RNA from HeLa cells. When we used Mono Q chromatography to investigate possible structural differences in the two types of U1 snRNPs, we observed that protein A was always preferentially lost from U1b snRNP as compared to U1a snRNPs. This indicates that one consequence of the structural difference between U1a and U1b is a lowering of the strength of binding of protein A to U1b snRNP. The possible functional significance of this finding is discussed with respect to the fact that U1b RNA is preferentially expressed in embryonal cells.

Citing Articles

Examining the capacity of human U1 snRNA variants to facilitate pre-mRNA splicing.

Wong J, Yellamaty R, Gallante C, Lawrence E, Martelly W, Sharma S RNA. 2024; 30(3):271-280.

PMID: 38164604 PMC: 10870369. DOI: 10.1261/rna.079892.123.

The regulation and pharmacological modulation of immune complex induced type III IFN production by plasmacytoid dendritic cells.

Hjorton K, Hagberg N, Pucholt P, Eloranta M, Ronnblom L Arthritis Res Ther. 2020; 22(1):130.

PMID: 32503683 PMC: 7275601. DOI: 10.1186/s13075-020-02186-z.

Developmental analysis of spliceosomal snRNA isoform expression.

Lu Z, Matera A G3 (Bethesda). 2014; 5(1):103-10.

PMID: 25416704 PMC: 4291461. DOI: 10.1534/g3.114.015735.

Structural analyses of the pre-mRNA splicing machinery.

Zhang L, Li X, Zhao R Protein Sci. 2013; 22(6):677-92.

PMID: 23592432 PMC: 3690710. DOI: 10.1002/pro.2266.

Limited complementarity between U1 snRNA and a retroviral 5' splice site permits its attenuation via RNA secondary structure.

Zychlinski D, Erkelenz S, Melhorn V, Baum C, Schaal H, Bohne J Nucleic Acids Res. 2009; 37(22):7429-40.

PMID: 19854941 PMC: 2794156. DOI: 10.1093/nar/gkp694.

References

Mattaj I, De Robertis E . Nuclear segregation of U2 snRNA requires binding of specific snRNP proteins. Cell. 1985; 40(1):111-8. DOI: 10.1016/0092-8674(85)90314-9. View

Billings P, Hoch S . Characterization of U small nuclear RNA-associated proteins. J Biol Chem. 1984; 259(20):12850-6. View

Zhuang Y, Weiner A . A compensatory base change in U1 snRNA suppresses a 5' splice site mutation. Cell. 1986; 46(6):827-35. DOI: 10.1016/0092-8674(86)90064-4. View

Mattaj I . Cap trimethylation of U snRNA is cytoplasmic and dependent on U snRNP protein binding. Cell. 1986; 46(6):905-11. DOI: 10.1016/0092-8674(86)90072-3. View

Aebi M, HORNIG H, Padgett R, Reiser J, Weissmann C . Sequence requirements for splicing of higher eukaryotic nuclear pre-mRNA. Cell. 1986; 47(4):555-65. DOI: 10.1016/0092-8674(86)90620-3. View

Sharp P . Splicing of messenger RNA precursors. Science. 1987; 235(4790):766-71. DOI: 10.1126/science.3544217. View

Theissen H, Etzerodt M, Reuter R, Schneider C, Lottspeich F, Argos P . Cloning of the human cDNA for the U1 RNA-associated 70K protein. EMBO J. 1986; 5(12):3209-17. PMC: 1167314. DOI: 10.1002/j.1460-2075.1986.tb04631.x. View

Maniatis T, REED R . The role of small nuclear ribonucleoprotein particles in pre-mRNA splicing. Nature. 1987; 325(6106):673-8. DOI: 10.1038/325673a0. View

Bringmann P, Luhrmann R . Purification of the individual snRNPs U1, U2, U5 and U4/U6 from HeLa cells and characterization of their protein constituents. EMBO J. 1986; 5(13):3509-16. PMC: 1167387. DOI: 10.1002/j.1460-2075.1986.tb04676.x. View

10.

Habets W, Sillekens P, Hoet M, Schalken J, Roebroek A, Leunissen J . Analysis of a cDNA clone expressing a human autoimmune antigen: full-length sequence of the U2 small nuclear RNA-associated B" antigen. Proc Natl Acad Sci U S A. 1987; 84(8):2421-5. PMC: 304663. DOI: 10.1073/pnas.84.8.2421. View

11.

Verheijen R, Kuijpers H, Vooijs P, Van Venrooij W, Ramaekers F . Distribution of the 70K U1 RNA-associated protein during interphase and mitosis. Correlation with other U RNP particles and proteins of the nuclear matrix. J Cell Sci. 1986; 86:173-90. DOI: 10.1242/jcs.86.1.173. View

12.

Query C, Keene J . A human autoimmune protein associated with U1 RNA contains a region of homology that is cross-reactive with retroviral p30gag antigen. Cell. 1987; 51(2):211-20. DOI: 10.1016/0092-8674(87)90148-6. View

13.

Bochnig P, Reuter R, Bringmann P, Luhrmann R . A monoclonal antibody against 2,2,7-trimethylguanosine that reacts with intact, class U, small nuclear ribonucleoproteins as well as with 7-methylguanosine-capped RNAs. Eur J Biochem. 1987; 168(2):461-7. DOI: 10.1111/j.1432-1033.1987.tb13439.x. View

14.

Spritz R, Strunk K, Surowy C, Hoch S, Barton D, Francke U . The human U1-70K snRNP protein: cDNA cloning, chromosomal localization, expression, alternative splicing and RNA-binding. Nucleic Acids Res. 1987; 15(24):10373-91. PMC: 339950. DOI: 10.1093/nar/15.24.10373. View

15.

Hamm J, Kazmaier M, Mattaj I . In vitro assembly of U1 snRNPs. EMBO J. 1987; 6(11):3479-85. PMC: 553806. DOI: 10.1002/j.1460-2075.1987.tb02672.x. View

16.

Sillekens P, Habets W, Beijer R, van Venrooij W . cDNA cloning of the human U1 snRNA-associated A protein: extensive homology between U1 and U2 snRNP-specific proteins. EMBO J. 1987; 6(12):3841-8. PMC: 553857. DOI: 10.1002/j.1460-2075.1987.tb02721.x. View

17.

Patton J, Pederson T . The Mr 70,000 protein of the U1 small nuclear ribonucleoprotein particle binds to the 5' stem-loop of U1 RNA and interacts with Sm domain proteins. Proc Natl Acad Sci U S A. 1988; 85(3):747-51. PMC: 279632. DOI: 10.1073/pnas.85.3.747. View

18.

Sillekens P, Beijer R, Habets W, van Venrooij W . Human U1 snRNP-specific C protein: complete cDNA and protein sequence and identification of a multigene family in mammals. Nucleic Acids Res. 1988; 16(17):8307-21. PMC: 338560. DOI: 10.1093/nar/16.17.8307. View

19.

Hamm J, van Santen V, Spritz R, Mattaj I . Loop I of U1 small nuclear RNA is the only essential RNA sequence for binding of specific U1 small nuclear ribonucleoprotein particle proteins. Mol Cell Biol. 1988; 8(11):4787-91. PMC: 365571. DOI: 10.1128/mcb.8.11.4787-4791.1988. View

20.

Mattaj I . A binding consensus: RNA-protein interactions in splicing, snRNPs, and sex. Cell. 1989; 57(1):1-3. DOI: 10.1016/0092-8674(89)90164-5. View