Functional characterization of the Tn5 transposase by limited proteolysis.

The Journal of Biological Chemistry
L Mahnke Braam, William S Reznikoff

Abstract

The 476 amino acid Tn5 transposase catalyzes DNA cutting and joining reactions that cleave the Tn5 transposon from donor DNA and integrate it into a target site. Protein-DNA and protein-protein interactions are important for this tranposition process. A truncated transposase variant, the inhibitor, decreases transposition rates via the formation of nonproductive complexes with transposase. Here, the inhibitor and the transposase are shown to have similar secondary and tertiary folding. Using limited proteolysis, the transposase has been examined structurally and functionally. A DNA binding region was localized to the N-terminal 113 amino acids. Generally, the N terminus of transposase is sensitive to proteolysis but can be protected by DNA. Two regions are predicted to contain determinants for protein-protein interactions, encompassing residues 114-314 and 441-476. The dimerization regions appear to be distinct and may have separate functions, one involved in synaptic complex formation and one involved in nonproductive multimerization. Furthermore, predicted catalytic regions are shown to lie between major areas of proteolysis.

References

Feb 1, 1992·Journal of Bacteriology·T W Wiegand, W S Reznikoff
Jul 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·A DeLong, M Syvanen
Jan 1, 1993·Annual Review of Microbiology·W S Reznikoff
Oct 20, 1995·Journal of Molecular Biology·G BujaczA M Skalka
Aug 29, 1995·Proceedings of the National Academy of Sciences of the United States of America·D KwonN Kleckner
Jan 1, 1995·Molecular Microbiology·P Polard, M Chandler
Sep 1, 1993·Molecular Microbiology·R RezsöhazyJ Mahillon
Nov 8, 1994·Proceedings of the National Academy of Sciences of the United States of America· Goryshin IYuW S Reznikoff
Jan 14, 1994·Journal of Molecular Biology·T W Wiegand, W S Reznikoff
Jul 1, 1993·Molecular Microbiology·C Jain, N Kleckner
Oct 1, 1996·Nucleic Acids Research·D York, W S Reznikoff
Aug 22, 1997·Journal of Molecular Biology·M Zhou, W S Reznikoff
Apr 29, 1998·The Journal of Biological Chemistry·I Y Goryshin, W S Reznikoff

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Citations

Dec 22, 1999·Biochemical and Biophysical Research Communications·W S ReznikoffS S Twining
Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·I Y GoryshinW S Reznikoff
Jul 5, 2005·Trends in Biotechnology·Craig J CoatesAndreas F Kolb
Mar 18, 2008·Insect Biochemistry and Molecular Biology·Jianjun WangYoonseong Park
Nov 5, 1999·Annual Review of Microbiology·L HarenM Chandler
Aug 31, 2000·Journal of Molecular Biology·A BhasinW S Reznikoff
May 30, 2006·The Journal of Biological Chemistry·Chang-Chieh Chen, Shiau-Ting Hu
Apr 17, 1999·The Journal of Biological Chemistry·D R DaviesI Rayment
Apr 3, 2001·The Journal of Biological Chemistry·S S TwiningW S Reznikoff
May 29, 2000·The Journal of Biological Chemistry·M Steiniger-White, W S Reznikoff
Dec 29, 1998·The Journal of Biological Chemistry·L A Mahnke BraamW S Reznikoff
Jun 25, 2015·Microbiology Spectrum·Alison B Hickman, Fred Dyda

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