Abstract
Photochemical cross-linking has been used to identify residues in the Escherichia coli RecA protein that are proximal to and may directly mediate binding of DNA. Ultraviolet irradiation promotes specific and efficient cross-linking of the RecA protein to poly(deoxythymidylic) acid. Cross- linked peptides remaining covalently attached to the polynucleotide following proteolytic digestion with trypsin correspond to amino acids 61-72, 178-183, and 233-243 of the RecA protein primary sequence. Their location and surface accessibility in the crystal structure, along with the behavior of various recA mutants, support the assignment of the cross-linked regions to the DNA binding site(s) of the RecA protein. Functional overlap of amino acids 61-72 with an element of the ATP binding site suggests a structural mechanism by which nucleotide cofactors allosterically affect the RecA nucleoprotein filament.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 11996-12002 |
| Number of pages | 7 |
| Journal | Journal of Biological Chemistry |
| Volume | 271 |
| Issue number | 20 |
| DOIs | |
| State | Published - 1996 |
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ASJC Scopus subject areas
- Biochemistry
Cite this
The DNA binding site(s) of the Escherichia coli RecA protein. / Rehrauer, William M.; Kowalczykowski, Stephen C.
In: Journal of Biological Chemistry, Vol. 271, No. 20, 1996, p. 11996-12002.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The DNA binding site(s) of the Escherichia coli RecA protein
AU - Rehrauer, William M.
AU - Kowalczykowski, Stephen C.
PY - 1996
Y1 - 1996
N2 - Photochemical cross-linking has been used to identify residues in the Escherichia coli RecA protein that are proximal to and may directly mediate binding of DNA. Ultraviolet irradiation promotes specific and efficient cross-linking of the RecA protein to poly(deoxythymidylic) acid. Cross- linked peptides remaining covalently attached to the polynucleotide following proteolytic digestion with trypsin correspond to amino acids 61-72, 178-183, and 233-243 of the RecA protein primary sequence. Their location and surface accessibility in the crystal structure, along with the behavior of various recA mutants, support the assignment of the cross-linked regions to the DNA binding site(s) of the RecA protein. Functional overlap of amino acids 61-72 with an element of the ATP binding site suggests a structural mechanism by which nucleotide cofactors allosterically affect the RecA nucleoprotein filament.
AB - Photochemical cross-linking has been used to identify residues in the Escherichia coli RecA protein that are proximal to and may directly mediate binding of DNA. Ultraviolet irradiation promotes specific and efficient cross-linking of the RecA protein to poly(deoxythymidylic) acid. Cross- linked peptides remaining covalently attached to the polynucleotide following proteolytic digestion with trypsin correspond to amino acids 61-72, 178-183, and 233-243 of the RecA protein primary sequence. Their location and surface accessibility in the crystal structure, along with the behavior of various recA mutants, support the assignment of the cross-linked regions to the DNA binding site(s) of the RecA protein. Functional overlap of amino acids 61-72 with an element of the ATP binding site suggests a structural mechanism by which nucleotide cofactors allosterically affect the RecA nucleoprotein filament.
UR - http://www.scopus.com/inward/record.url?scp=17544384169&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=17544384169&partnerID=8YFLogxK
U2 - 10.1074/jbc.271.20.11996
DO - 10.1074/jbc.271.20.11996
M3 - Article
C2 - 8662640
AN - SCOPUS:17544384169
VL - 271
SP - 11996
EP - 12002
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 20
ER -