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PMID: 9271394 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Characterization of the maize Mutator transposable element MURA transposase as a DNA-binding protein.

Molecular and cellular biology ·Vol. 17 ·No. 9 ·1997-09-00 ·Pages 5165-75

Benito MI, Walbot V

Abstract

The autonomous MuDR element of the Mutator (Mu) transposable element family of maize encodes at least two proteins, MURA and MURB. Based on amino acid sequence similarity, previous studies have reported that MURA is likely to be a transposase. The functional characterization of MURA has been hindered by the instability of its cDNA, mudrA, in Escherichia coli. In this study, we report the first successful stabilization and expression of MURA in Saccharomyces cerevisiae. Gel mobility shift assays demonstrate that MURA is a DNA-binding protein that specifically binds to sequences within the highly conserved Mu element terminal inverted repeats (TIRs). DNase I and 1,10-phenanthroline-copper footprinting of MURA-Mu1 TIR complexes indicate that MURA binds to a conserved approximately 32-bp region in the TIR of Mu1. In addition, MURA can bind to the same region in the TIRs of all tested actively transposing Mu elements but binds poorly to the diverged Mu TIRs of inactive elements. Previous studies have reported a correlation between Mu transposon inactivation and methylation of the Mu element TIRs. Gel mobility shift assays demonstrate that MURA can interact differentially with unmethylated, hemimethylated, and homomethylated TIR substrates. The significance of MURA's interaction with the TIRs of Mu elements is discussed in the context of what is known about the regulation and mechanisms of Mutator activities in maize.

MeSH Terms
Base Sequence Copper/metabolism DNA Footprinting DNA Methylation DNA Nucleotidyltransferases/chemistry,metabolism DNA Transposable Elements DNA-Binding Proteins/chemistry,metabolism Molecular Sequence Data Phenanthrolines/metabolism Protein Binding Saccharomyces cerevisiae Transposases Zea mays/enzymology,genetics
Chemicals
DNA Transposable Elements DNA-Binding Proteins Phenanthrolines Copper DNA Nucleotidyltransferases Transposases 1,10-phenanthroline
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Benito M I
Department of Biological Sciences, Stanford University, California 94305-5020, USA. [email protected]
Walbot V
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.9.5165
Email
[email protected]
Published
1997-09-00
Pages
5165-75
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232367
Subset
IM
Grants
NIGMS NIH HHS · GM49681 · United States
External Links
PubMed source DOI: 10.1128/MCB.17.9.5165 PMC Full Text View journal details
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