This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jamison, S F Articles by Garcia-Blanco, M A Search for Related Content PubMed PubMed Citation Articles by Jamison, S F Articles by Garcia-Blanco, M A

 Previous Article  |  Next Article 

Mol Cell Biol. 1992 October; 12(10): 4279-4287

The spliceosome assembly pathway in mammalian extracts.

Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina 27710.

ABSTRACT

A mammalian splicing commitment complex was functionally defined by using a template commitment assay. This complex was partially purified and shown to be a required intermediate for complex A formation. The productive formation of this commitment complex required both splice sites and the polypyrimidine tract. U1 small nuclear ribonucleoprotein (snRNP) was the only spliceosomal U snRNP required for this formation. A protein factor, very likely U2AF, is probably involved in the formation of the splicing commitment complex. From the kinetics of appearance of complex A and complex B, it was previously postulated that complex A represents a functional intermediate in spliceosome assembly. Complex A was partially purified and shown to be a required intermediate for complex B (spliceosome) formation. Thus, a spliceosome pathway is for the first time supported by direct biochemical evidence: RNA+U1 snRNP+?U2 auxiliary factor+?Y----CC+U2 snRNP+Z----A+U4/6,5 snRNPs+ beta----B.

This article has been cited by other articles:

• Zhou, H.-L., Lou, H. (2008). Repression of Prespliceosome Complex Formation at Two Distinct Steps by Fox-1/Fox-2 Proteins. Mol. Cell. Biol. 28: 5507-5516 [Abstract] [Full Text]  
• Frilander, M. J., Meng, X. (2005). Proximity of the U12 snRNA with both the 5' Splice Site and the Branch Point during Early Stages of Spliceosome Assembly. Mol. Cell. Biol. 25: 4813-4825 [Abstract] [Full Text]  
• Kent, O. A., Ritchie, D. B., MacMillan, A. M. (2005). Characterization of a U2AF-Independent Commitment Complex (E') in the Mammalian Spliceosome Assembly Pathway. Mol. Cell. Biol. 25: 233-240 [Abstract] [Full Text]  
• (2001). Prespliceosomal Assembly on Microinjected Precursor mRNA Takes Place in Nuclear Speckles. Mol. Biol. Cell 12: 393-406 [Abstract] [Full Text]  
• Zhang, M., Green, M. R. (2001). Identification and characterization of yUAP/Sub2p, a yeast homolog of the essential human pre-mRNA splicing factor hUAP56. Genes Dev. 15: 30-35 [Abstract] [Full Text]  
• Frilander, M. J., Steitz, J. A. (1999). Initial recognition of U12-dependent introns requires both U11/5' splice-site and U12/branchpoint interactions. Genes Dev. 13: 851-863 [Abstract] [Full Text]  
• Labourier, E., Bourbon, H.-M., Gallouzi, I.-e., Fostier, M., Allemand, E., Tazi, J. (1999). Antagonism between RSF1 and SR proteins for both splice-site recognition in vitro and Drosophila development. Genes Dev. 13: 740-753 [Abstract] [Full Text]  
• Liu, Z.-R., Sargueil, B., Smith, C. W. J. (1998). Detection of a Novel ATP-Dependent Cross-Linked Protein at the 5' Splice Site-U1 Small Nuclear RNA Duplex by Methylene Blue-Mediated Photo-Cross-Linking. Mol. Cell. Biol. 18: 6910-6920 [Abstract] [Full Text]  
• Zheng, Z. M., Huynen, M., Baker, C. C. (1998). A pyrimidine-rich exonic splicing suppressor binds multiple RNA splicing factors and inhibits spliceosome assembly. Proc. Natl. Acad. Sci. USA 95: 14088-14093 [Abstract] [Full Text]  
• Cao, W., Garcia-Blanco, M. A. (1998). A Serine/Arginine-rich Domain in the Human U1 70k Protein Is Necessary and Sufficient for ASF/SF2 Binding. J. Biol. Chem. 273: 20629-20635 [Abstract] [Full Text]  
• Bouck, J., Fu, X.-D., Skalka, A. M., Katz, R. A. (1998). Role of the Constitutive Splicing Factors U2AF65 and SAP49 in Suboptimal RNA Splicing of Novel Retroviral Mutants. J. Biol. Chem. 273: 15169-15176 [Abstract] [Full Text]  
• Askjaer, P., Kjems, J. (1998). Mapping of Multiple RNA Binding Sites of Human T-cell Lymphotropic Virus Type I Rex Protein within 5'- and 3'-Rex Response Elements. J. Biol. Chem. 273: 11463-11471 [Abstract] [Full Text]  
• Ruby, S. W. (1997). Dynamics of the U1 Small Nuclear Ribonucleoprotein during Yeast Spliceosome Assembly. J. Biol. Chem. 272: 17333-17341 [Abstract] [Full Text]  
• Xiao, S H, Manley, J L (1997). Phosphorylation of the ASF/SF2 RS domain affects both protein-protein and protein-RNA interactions and is necessary for splicing.. Genes Dev. 11: 334-344 [Abstract]  
• Dyhr-Mikkelsen, H., Kjems, Jør. (1995). Inefficient Spliceosome Assembly and Abnormal Branch Site Selection in Splicing of an HIV-1 Transcript in Vitro. J. Biol. Chem. 270: 24060-24066 [Abstract] [Full Text]  
• Furman, E., Glitz, D. G. (1995). Purification of the Spliceosome A-Complex and Its Visualization by Electron Microscopy. J. Biol. Chem. 270: 15515-15522 [Abstract] [Full Text]  
• Lindsey, L. A., Crow, A. J., Garcia-Blanco, M. A. (1995). A Mammalian Activity Required for the Second Step of Pre-messenger RNA Splicing. J. Biol. Chem. 270: 13415-13421 [Abstract] [Full Text]  
• Singh, R, Valcarcel, J, Green, M. (1995). Distinct binding specificities and functions of higher eukaryotic polypyrimidine tract-binding proteins. Science 268: 1173-1176 [Abstract]  
• Berget, S. M., Berget, S. M. (1995). Exon Recognition in Vertebrate Splicing. J. Biol. Chem. 270: 2411-2414 [Full Text]  
• Abovich, N, Liao, X C, Rosbash, M (1994). The yeast MUD2 protein: an interaction with PRP11 defines a bridge between commitment complexes and U2 snRNP addition.. Genes Dev. 8: 843-854 [Abstract]  
• Norton, P. (1994). Alternative pre-mRNA splicing: factors involved in splice site selection. J. Cell Sci. 107: 1-7  
• Lesser, C., Guthrie, C (1993). Mutations in U6 snRNA that alter splice site specificity: implications for the active site. Science 262: 1982-1988 [Abstract]  
• Michaud, S, Reed, R (1993). A functional association between the 5' and 3' splice site is established in the earliest prespliceosome complex (E) in mammals.. Genes Dev. 7: 1008-1020 [Abstract]  
• Patton, J G, Porro, E B, Galceran, J, Tempst, P, Nadal-Ginard, B (1993). Cloning and characterization of PSF, a novel pre-mRNA splicing factor.. Genes Dev. 7: 393-406 [Abstract]  
• Liao, X C, Tang, J, Rosbash, M (1993). An enhancer screen identifies a gene that encodes the yeast U1 snRNP A protein: implications for snRNP protein function in pre-mRNA splicing.. Genes Dev. 7: 419-428 [Abstract]  
• Blencowe, B., Carmo-Fonseca, M, Behrens, S., Luhrmann, R, Lamond, A. (1993). Interaction of the human autoantigen p150 with splicing snRNPs. J. Cell Sci. 105: 685-697 [Abstract]  
• Rappsilber, J., Ajuh, P., Lamond, A. I., Mann, M. (2001). SPF30 Is an Essential Human Splicing Factor Required for Assembly of the U4/U5/U6 Tri-small Nuclear Ribonucleoprotein into the Spliceosome. J. Biol. Chem. 276: 31142-31150 [Abstract] [Full Text]