U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition

Characterization of required elements

Sharon F. Jamison, Zvi Pasman, Jin Wang, Cindy Will, Reinhard Lührmann, James L. Manley, Mariano Garcia-Blanco

Research output: Contribution to journalArticle

95 Citations (Scopus)

Abstract

Members of the SR family of proteins, can collaborate with U1 snRNP in the recognition of 5′ splice sites in pre-messenger RNAs. We have previously shown that purified U1 snRNP and ASF/SF2 form a ternary complex with pre-mRNA, which is dependent on a functional 5′ splice site. In this manuscript we dissect the requirements for the formation of this complex. Sequences in the pre-mRNA, domains in ASF/SF2 and components of the U1 snRNP particle are shown to be required for complex formation. We had shown that sequences at the 5′ splice site of PIP7.A are necessary and now we show these are sufficient for complex formation. Furthermore, we show that one functional RNA binding domain and the RS domain are both required for ASF/SF2 to participate in complex formation. The RNA binding domains were redundant in this assay, suggesting that either domain can interact with the pre-messenger RNA. Finally, our experiments show no function for the U1-specific A protein in complex formation, whereas a function for U1-specific C protein was strongly suggested. The study of the earliest interactions between pre-mRNA and splicing factors suggests a model for 5′ splice site recognition.

Original languageEnglish (US)
Pages (from-to)3260-3267
Number of pages8
JournalNucleic Acids Research
Volume23
Issue number16
StatePublished - Aug 25 1995
Externally publishedYes

Fingerprint

U1 Small Nuclear Ribonucleoproteins
RNA Splice Sites
RNA Precursors
Messenger RNA
Interaction
Proteins
RNA
Protein
Protein C
Dissect
Assays
Ternary
Sufficient
Necessary
Dependent
Experiments
Requirements

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Jamison, S. F., Pasman, Z., Wang, J., Will, C., Lührmann, R., Manley, J. L., & Garcia-Blanco, M. (1995). U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition: Characterization of required elements. Nucleic Acids Research, 23(16), 3260-3267.

U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition : Characterization of required elements. / Jamison, Sharon F.; Pasman, Zvi; Wang, Jin; Will, Cindy; Lührmann, Reinhard; Manley, James L.; Garcia-Blanco, Mariano.

In: Nucleic Acids Research, Vol. 23, No. 16, 25.08.1995, p. 3260-3267.

Research output: Contribution to journalArticle

Jamison, SF, Pasman, Z, Wang, J, Will, C, Lührmann, R, Manley, JL & Garcia-Blanco, M 1995, 'U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition: Characterization of required elements', Nucleic Acids Research, vol. 23, no. 16, pp. 3260-3267.
Jamison SF, Pasman Z, Wang J, Will C, Lührmann R, Manley JL et al. U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition: Characterization of required elements. Nucleic Acids Research. 1995 Aug 25;23(16):3260-3267.
Jamison, Sharon F. ; Pasman, Zvi ; Wang, Jin ; Will, Cindy ; Lührmann, Reinhard ; Manley, James L. ; Garcia-Blanco, Mariano. / U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition : Characterization of required elements. In: Nucleic Acids Research. 1995 ; Vol. 23, No. 16. pp. 3260-3267.
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