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Cis preference of the IS 903 transposase is mediated by a combination of transposase instability and inefficient translation
Author(s) -
Derbyshire Keith M.,
Grindley Nigel D. F.
Publication year - 1996
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1996.tb02587.x
Subject(s) - transposase , biology , preference , translation (biology) , instability , genetics , transposable element , messenger rna , gene , physics , microeconomics , economics , mechanics , genome
Summary The transposase protein encoded by the insertion element IS 903 belongs to an unusual class of DNA‐binding proteins, termed cis ‐acting proteins, that act preferentially at their site of synthesis. Previous work had led us to propose that instability of the IS 903 transposase was a major determinant of its cis preference. Here we describe the isolation of two classes of mutations within the transposase gene that increased action in trans. One class specifically increased trans action without increasing the level of transposition when the mutant gene was located in cis to the transposon. In particular, a threonine‐to‐proline substitution at amino acid 25 (T25P) reduced cis preference about 60‐fold. The half‐life of this mutant transposase was significantly longer than that of the wild‐type transposase, confirming the critical role of protein instability. The second, larger, class of mutations increased the level of transposition both in trans and in cis . The behaviour and location of these mutations were consistent with an increase in gene expression by improving translational initiation. Several of these mutations exerted a disproportionate effect on the action of transposase in trans , implying that translation efficiency may affect more than just the amount of transposase made. Our results indicate that cis preference of the IS 903 transposase is mediated by a combination of transposase instability and inefficient translation initiation.