Nucleic Acids Research, Vol 25, Issue 11 2146-2152, Copyright © 1997 by Oxford University Press
X Zhang and B Schwer
We show that the requirement for Prp18 during the second step of actin
pre-mRNA splicing in vitro is dictated by the distance between the branch
point and the 3'splice site. Prp18 is dispensable for splicing of precursor
RNAs in which the interval between the branch point and 3'splice site is
<12 nt. This resembles the requirement for another second step factor,
Slu7. Excess Slu7 protein can bypass the need for Prp18 in vitro ,
suggesting that Slu7 and Prp18 function in a concerted manner. Physical
interaction between Slu7 and Prp18 was demonstrated by using the two-hybrid
assay. Deletion mutants of SLU7 were tested for their ability to support
growth of a slu7 null strain. Removal of 199 amino acids from the
N-terminus of the 382 amino acid Slu7 protein did not affect cell viability
at 25 degrees C. A more extensive N-terminal deletion of 221 amino acids
was lethal, as was a C-terminal deletion of 47 amino acids. Deleted
versions of Slu7 were also tested for interaction with Prp18 in the
two-hybrid system. We define a segment of Slu7 from residue 200 to 224 that
is necessary for interaction with Prp18.
ARTICLES
Functional and physical interaction between the yeast splicing factors Slu7 and Prp18
Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
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