» Articles » PMID: 17579712

Control of Pre-mRNA Splicing by the General Splicing Factors PUF60 and U2AF(65)

Overview
Journal PLoS One
Date 2007 Jun 21
PMID 17579712
Citations 70
Authors
Affiliations
Soon will be listed here.
Abstract

Pre-mRNA splicing is a crucial step in gene expression, and accurate recognition of splice sites is an essential part of this process. Splice sites with weak matches to the consensus sequences are common, though it is not clear how such sites are efficiently utilized. Using an in vitro splicing-complementation approach, we identified PUF60 as a factor that promotes splicing of an intron with a weak 3' splice-site. PUF60 has homology to U2AF(65), a general splicing factor that facilitates 3' splice-site recognition at the early stages of spliceosome assembly. We demonstrate that PUF60 can functionally substitute for U2AF(65)in vitro, but splicing is strongly stimulated by the presence of both proteins. Reduction of either PUF60 or U2AF(65) in cells alters the splicing pattern of endogenous transcripts, consistent with the idea that regulation of PUF60 and U2AF(65) levels can dictate alternative splicing patterns. Our results indicate that recognition of 3' splice sites involves different U2AF-like molecules, and that modulation of these general splicing factors can have profound effects on splicing.

Citing Articles

PUF60 Promotes Chemoresistance Through Drug Efflux and Reducing Apoptosis in Gastric Cancer.

Liu Q, Song Y, Su J, Yang S, Lian Q, Wang T Int J Med Sci. 2025; 22(2):269-282.

PMID: 39781520 PMC: 11704696. DOI: 10.7150/ijms.102976.


Nuclear translocation of SIRT4 mediates deacetylation of U2AF2 to modulate renal fibrosis through alternative splicing-mediated upregulation of CCN2.

Yang G, Xiang J, Yang X, Liu X, Li Y, Li L Elife. 2024; 13.

PMID: 39495216 PMC: 11534337. DOI: 10.7554/eLife.98524.


Recurrent Neurodevelopmentally Associated Variants of the Pre-mRNA Splicing Factor U2AF2 Alter RNA Binding Affinities and Interactions.

Maji D, Jenkins J, Boutz P, Kielkopf C Biochemistry. 2024; 63(21):2718-2722.

PMID: 39388459 PMC: 11542177. DOI: 10.1021/acs.biochem.4c00344.


Branch site recognition by the spliceosome.

Tholen J RNA. 2024; 30(11):1397-1407.

PMID: 39187383 PMC: 11482624. DOI: 10.1261/rna.080198.124.


Novel Genetic and Phenotypic Expansion in Ameliorated -Related Disorders.

Baum E, Huang W, Vincent-Delorme C, Brunelle P, Antebi A, Dafsari H Int J Mol Sci. 2024; 25(4).

PMID: 38396730 PMC: 10889399. DOI: 10.3390/ijms25042053.


References
1.
Sakamoto H, Ohno M, Yasuda K, Mizumoto K, Shimura Y . In vitro splicing of a chicken delta-crystallin pre-mRNA in a mammalian nuclear extract. J Biochem. 1987; 102(5):1289-301. DOI: 10.1093/oxfordjournals.jbchem.a122167. View

2.
Zamore P, Green M . Identification, purification, and biochemical characterization of U2 small nuclear ribonucleoprotein auxiliary factor. Proc Natl Acad Sci U S A. 1989; 86(23):9243-7. PMC: 298470. DOI: 10.1073/pnas.86.23.9243. View

3.
Tress M, Martelli P, Frankish A, Reeves G, Wesselink J, Yeats C . The implications of alternative splicing in the ENCODE protein complement. Proc Natl Acad Sci U S A. 2007; 104(13):5495-500. PMC: 1838448. DOI: 10.1073/pnas.0700800104. View

4.
Mayeda A, Krainer A . Preparation of HeLa cell nuclear and cytosolic S100 extracts for in vitro splicing. Methods Mol Biol. 1999; 118:309-14. DOI: 10.1385/1-59259-676-2:309. View

5.
MacMillan A, McCaw P, Crispino J, Sharp P . SC35-mediated reconstitution of splicing in U2AF-depleted nuclear extract. Proc Natl Acad Sci U S A. 1997; 94(1):133-6. PMC: 19256. DOI: 10.1073/pnas.94.1.133. View