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The 100-kDa U5 snRNP protein (hPrp28p) contacts the 5′ splice site through its ATPase site

Published online by Cambridge University Press:  07 February 2001

NAÏMA ISMAÏLI ,
MA SHA ,
E. HILARY GUSTAFSON  and
MARIA M. KONARSKA
NAÏMA ISMAÏLI
Affiliation:
The Rockefeller University, New York, New York 10021, USA
MA SHA
Affiliation:
The Rockefeller University, New York, New York 10021, USA
E. HILARY GUSTAFSON
Affiliation:
The Rockefeller University, New York, New York 10021, USA
MARIA M. KONARSKA
Affiliation:
The Rockefeller University, New York, New York 10021, USA
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Abstract

To identify splicing factors in proximity of the 5′ splice site (5′SS), we followed a crosslinking profile of site-specifically modified, photoreactive RNA substrates. Upon U4/U5/U6 snRNP addition, the 5′SS RNA crosslinks in an ATP-dependent manner to U6 snRNA, an unidentified protein p27, and the 100-kDa U5 snRNP protein, a human ortholog of an ATPase/RNA helicase yPrp28p. The 5′SS:hPrp28p crosslink maps to the highly conserved TAT motif in proximity of the ATP-binding site in hPrp28p. We propose that hPrp28p acts as a helicase to unwind the 5′SS:U1 snRNA duplex, and at the same time as a 5′SS translocase, which, upon NTP-dependent conformational change, positions the 5′SS for pairing with U6 snRNA within the spliceosome. This repositioning of the 5′SS takes place regardless of whether the 5′SS is originally duplexed with U1 snRNA.

Keywords

5′ splice sitepre-mRNA splicingPrp28RNA helicasespliceosome
Type
Research Article
Information
RNA , Volume 7 , Issue 2 , February 2001 , pp. 182 - 193
Copyright
2001 RNA Society

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