TY - JOUR
T1 - U1 snRNP-ASF/SF2 interaction and 5′ splice site recognition
T2 - Characterization of required elements
AU - Jamison, Sharon F.
AU - Pasman, Zvi
AU - Wang, Jin
AU - Will, Cindy
AU - Lührmann, Reinhard
AU - Manley, James L.
AU - Garcia-blanco, Mariano A.
N1 - Funding Information:
We thank J. Keene (Duke University) for much encouragement and for providing us with Sm antisera. We are grateful to Sabina Sager for assistance in the preparation of this manuscript. This work was funded by research grants from NTH (M.A.G.-B. and J.L.M). M.A.G.-B. is funded by a JFRA from the ACS and would also like to acknowledge the support of the Keck Foundation to the Levine Science Research Center.
PY - 1995/8/25
Y1 - 1995/8/25
N2 - 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.
AB - 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.
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U2 - 10.1093/nar/23.16.3260
DO - 10.1093/nar/23.16.3260
M3 - Article
C2 - 7667103
AN - SCOPUS:0029090081
SN - 0305-1048
VL - 23
SP - 3260
EP - 3267
JO - Nucleic acids research
JF - Nucleic acids research
IS - 16
ER -