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Structural Domains in Phage Mu Transposase: Identification of the Site-Specific DNA-Binding Domain

Nakayama, Chikao and Teplow, David B. and Harshey, Rasika M. (1987) Structural Domains in Phage Mu Transposase: Identification of the Site-Specific DNA-Binding Domain. Proceedings of the National Academy of Sciences of the United States of America, 84 (7). pp. 1809-1813. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:NAKpnas87

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Abstract

Limited proteolysis of phage Mu transposase with three proteases of differing specificities produced a common pattern of fragmentation. The fragments were mapped by using a combination of immunoblotting and amino acid sequence analysis. Our results suggest that the transposase molecule is organized principally into three domains: an amino-terminal domain of molecular mass 30 kDa, a core region of approximately 35 kDa, and a carboxyl-terminal domain of approximately 10 kDa. The amino-terminal domain has at least two additional sites that are partially accessible to proteases. Filter binding and nuclease protection studies were done to determine the functions of the isolated domains. Site-specific binding to Mu DNA was localized to the amino-terminal domain. The core domain showed nonspecific DNA-binding activity.


Item Type:Article
Additional Information:Copyright © 1987 by the National Academy of Sciences Communicated by Leroy E. Hood, November 19, 1986 We thank R. Houghten for peptide synthesis, Stephen Kent and Lee Hood for encouragement and support, Ruedi Aebersold for helpful discussions, and Connie Katz and Cathy Elkins for expert secretarial assistance. This work was supported by National Institutes of Health Grant GM 33247 (R.M.H.) and by National Science Foundation Grant DMB 85-00298 (D.B.T.). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:transposable elements; gene rearrangements; limited proteolysis; protein structure; protein-DNA interaction
Record Number:CaltechAUTHORS:NAKpnas87
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:NAKpnas87
Alternative URL:http://www.pnas.org/cgi/content/abstract/84/7/1809
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5790
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:02 Nov 2006
Last Modified:26 Dec 2012 09:15

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