An Intronic Splicing Enhancer Binds U1 snRNPs To Enhance Splicing and Select 5' Splice Sites

doi:10.1128/MCB.20.24.9225-9235.2000

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Molecular and Cellular Biology, December 2000, p. 9225-9235, Vol. 20, No. 24
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

An Intronic Splicing Enhancer Binds U1 snRNPs To Enhance Splicing and Select 5' Splice Sites

Andrew J. McCullough^* and Susan M. Berget

Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030

Received 19 April 2000/Returned for modification 7 June 2000/Accepted 19 September 2000

Intronic G triplets are frequently located adjacent to 5' splice sites in vertebrate pre-mRNAs and have been correlated withsplicing efficiency and specificity via a mechanism that activatesupstream 5' splice sites in exons containing duplicated sites(26). Using an intron dependent upon G triplets for maximal activityand 5' splice site specificity, we determined that these elementsbind U1 snRNPs via base pairing with U1 RNA. This interactionis novel in that it uses nucleotides 8 to 10 of U1 RNA and isindependent of nucleotides 1 to 7. In vivo functionality of basepairing was documented by restoring activity and specificity tomutated G triplets through compensating U1 RNA mutations. We suggestthat the G-rich region near vertebrate 5' splice sites promotesaccurate splice site recognition by recruiting the U1snRNP.

^* Corresponding author. Mailing address: Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston,TX 77030. Phone: (713) 798-4622. Fax: (713) 795-5487. E-mail:andrewm{at}bcm.tmc.edu.

Molecular and Cellular Biology, December 2000, p. 9225-9235, Vol. 20, No. 24
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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