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osu1254972783.pdf (9.11 MB)
ETD Abstract Container
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Structural Studies of Escherichia coli RecE Exonuclease
Author Info
Zhang, Jinjin
Permalink:
http://rave.ohiolink.edu/etdc/view?acc_num=osu1254972783
Abstract Details
Year and Degree
2009, Doctor of Philosophy, Ohio State University, Ohio State Biochemistry Program.
Abstract
E. coli RecE protein is part of the classical RecET recombination system that has recently been employed in powerful new methods for genetic engineering. RecE binds to free dsDNA ends and processively digests the 5’-ended strand to form 5’-mononucleotides and a 3’-overhang that is a substrate for single strand annealing promoted by RecT. Here, we report the crystal structure of the C-terminal nuclease domain of RecE at 2.8 Å resolution. RecE forms a toroidal tetramer with a central tapered channel that is wide enough to bind dsDNA at one end, but is partially plugged at the other end by the C-terminal segment of the protein. Four narrow tunnels, one within each subunit of the tetramer, lead from the central channel to the four active sites, which lie about 15 Å from the channel. The structure, combined with mutational studies, suggests a mechanism in which dsDNA enters through the open end of the central channel, the 5’-ended strand passes through a tunnel to access one of the four active sites, and the 3’-ended strand passes through the plugged end of the channel at the back of the tetramer.
Committee
Charles Bell, PhD (Advisor)
Ross Dalbey, PhD (Committee Member)
Dehua Pei, PhD (Committee Member)
Jiyan Ma, PhD (Committee Member)
Pages
160 p.
Subject Headings
Biochemistry
Keywords
RecE
;
exonuclease
;
crystal structure
;
DNA binding
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Citations
Zhang, J. (2009).
Structural Studies of Escherichia coli RecE Exonuclease
[Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1254972783
APA Style (7th edition)
Zhang, Jinjin.
Structural Studies of Escherichia coli RecE Exonuclease.
2009. Ohio State University, Doctoral dissertation.
OhioLINK Electronic Theses and Dissertations Center
, http://rave.ohiolink.edu/etdc/view?acc_num=osu1254972783.
MLA Style (8th edition)
Zhang, Jinjin. "Structural Studies of Escherichia coli RecE Exonuclease." Doctoral dissertation, Ohio State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=osu1254972783
Chicago Manual of Style (17th edition)
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Document number:
osu1254972783
Download Count:
579
Copyright Info
© 2009, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.