Organization and dynamics of the bacteriophage Mu transpososome

Transposition of a linear mobile genetic element, such as the bacteriophage Mu, requires a set of spatially and temporally coordinated DNA phosphoryl transfer reactions to move the element from an existing DNA location to a new DNA site. The Mu transpososome, where the DNA cleavage and joining reactions that compose transposition take place, is a large nucleoprotein complex containing multiple, identical transposase subunits. The cleavage and joining reactions are the result of precise manipulation, by the transposase subunits, of the DNA segments surrounding the two transposon ends and the DNA at the new location. The purpose of this work is to provide a description of the arrangement of the catalytic centers within the Mu transpososome and of the dynamic interaction of the DNA components with the active sites. By employing a unique protein-DNA crosslinking strategy developed in this thesis work, the transposase subunits that contribute three essential acidic amino acids (the DDE motif) to the active sites are identified. This work reveals that the transpososome contains two active sites, as defined by the number of required sets of DDE residues, each of which catalyzes cleavage and joining of one transposon DNA end to the new location. The organization of the subunits supplying these active site residues is such that a subunit bound site-specifically to one transposon DNA end contributes DDE residues to the active site that promotes the phosphoryl transfer reactions at the other transposon end.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Chemistry.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2002

Author dc:creator

• Williams, Tanya L. (Tanya Lynn), 1970-

Advisor dc:contributor.advisor

• Tania A. Baker.

• Chemistry.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/8364

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/8364