Mutations in the mariner transposase: the D,D(35)E consensus sequence is nonfunctional

Citation:

Lohe, AR, D De Aguiar, and DL Hartl. 1997. “Mutations in the mariner transposase: the D,D(35)E consensus sequence is nonfunctional.” Proc Natl Acad Sci U S A 94: 1293-7.

Date Published:

Feb 18

Abstract:

Genetic analysis of eukaryote transposases and comparison with their prokaryote counterparts have been greatly hindered by difficulty in isolating mutations. We describe a simple eye-color screen that facilitates isolation and analysis of mutations in the mariner transposase in Drosophila melanogaster. Use of ethyl methanesulfonate and site-directed mutagenesis has identified 18 residues that are critical for in vivo excision of a target mariner element. When the mutations were examined in heterozygous mutant/nonmutant genotypes, more than half of the mutant transposase proteins were found to reduce the activity of the wild-type transposase, as assayed by the frequency of germline excision of a target element. Remarkably, transposase function is obliterated when the D,D(34)D acidic, ion-binding domain is replaced with the consensus sequence D,D(34)E found in the nematode Tc1 transposase and in many other transposases in the superfamily. A number of mutations strongly complement wild-type transposase in a dominant-negative manner, suggestive of subunit interactions in the excision reaction; these mutations are located in a small region that includes part of the D,D(34)D motif. Transposase function also is eliminated by a mutation in the inferred initiation codon and by a mutation in a putative nuclear localization signal.

Notes:

Lohe, A RDe Aguiar, DHartl, D LengGM33741/GM/NIGMS NIH HHS/Research Support, U.S. Gov't, P.H.S.1997/02/18Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1293-7.

Last updated on 05/20/2015