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ATP requirement for Prp5p function is determined by Cus2p and the structure of U2 small nuclear RNA
 

Summary: ATP requirement for Prp5p function is determined by
Cus2p and the structure of U2 small nuclear RNA
Rhonda Perriman*, Imre Barta
, Gia K. Voeltz*
, John Abelson
, and Manuel Ares, Jr.*
*Center for Molecular Biology of RNA, Sinsheimer Laboratories, University of California, Santa Cruz, CA 95064; and Division of Biology, California Institute
of Technology, Pasadena, CA 91125
Contributed by John Abelson, September 30, 2003
Stable addition of U2 small nuclear ribonucleoprotein (snRNP) to
form the prespliceosome is the first ATP-dependent step in splicing,
and it requires the DEXD H box ATPase Prp5p. However, prespli-
ceosome formation occurs without ATP in extracts lacking the U2
snRNP protein Cus2p. Here we show that Prp5p is required for the
ATP-independent prespliceosome assembly that occurs in the ab-
sence of Cus2p. Addition of recombinant Cus2p can restore the ATP
dependence of prespliceosome assembly, but only if it is added
before Prp5p. Prp5p with an altered ATP-binding domain (Prp5-
GNTp) can support growth in vivo, but only in a cus2 deletion
strain, mirroring the in vitro results. Other Prp5 ATP-binding do-

  

Source: Ares Jr., Manny - Department of Molecular, Cell, and Developmental Biology, University of California at Santa Cruz

 

Collections: Biology and Medicine