dc.contributor.author |
Augustus, Anne M |
|
dc.contributor.author |
Sage, Harvey |
|
dc.contributor.author |
Spicer, Leonard D |
|
dc.coverage.spatial |
United States |
|
dc.date.accessioned |
2011-06-21T17:22:11Z |
|
dc.date.issued |
2010-04-20 |
|
dc.identifier |
http://www.ncbi.nlm.nih.gov/pubmed/20196619 |
|
dc.identifier.uri |
https://hdl.handle.net/10161/4018 |
|
dc.description.abstract |
We have used analytical ultracentrifugation to characterize the binding of the methionine
repressor protein, MetJ, to synthetic oligonucleotides containing zero to five specific
recognition sites, called metboxes. For all lengths of DNA studied, MetJ binds more
tightly to repeats of the consensus sequence than to naturally occurring metboxes,
which exhibit a variable number of deviations from the consensus. Strong cooperative
binding occurs only in the presence of two or more tandem metboxes, which facilitate
protein-protein contacts between adjacent MetJ dimers, but weak affinity is detected
even with DNA containing zero or one metbox. The affinity of MetJ for all of the DNA
sequences studied is enhanced by the addition of SAM, the known cofactor for MetJ
in the cell. This effect extends to oligos containing zero or one metbox, both of
which bind two MetJ dimers. In the presence of a large excess concentration of metbox
DNA, the effect of cooperativity is to favor populations of DNA oligos bound by two
or more MetJ dimers rather than a stochastic redistribution of the repressor onto
all available metboxes. These results illustrate the dynamic range of binding affinity
and repressor assembly that MetJ can exhibit with DNA and the effect of the corepressor
SAM on binding to both specific and nonspecific DNA.
|
|
dc.language |
eng |
|
dc.language.iso |
en_US |
|
dc.publisher |
American Chemical Society (ACS) |
|
dc.relation.ispartof |
Biochemistry |
|
dc.relation.isversionof |
10.1021/bi902011f |
|
dc.subject |
Bacterial Proteins |
|
dc.subject |
Base Sequence |
|
dc.subject |
Consensus Sequence |
|
dc.subject |
DNA |
|
dc.subject |
Dimerization |
|
dc.subject |
Fractionation, Field Flow |
|
dc.subject |
Kinetics |
|
dc.subject |
Methionine |
|
dc.subject |
Molecular Weight |
|
dc.subject |
Oligodeoxyribonucleotides |
|
dc.subject |
Protein Binding |
|
dc.subject |
Repressor Proteins |
|
dc.subject |
S-Adenosylmethionine |
|
dc.subject |
Ultracentrifugation |
|
dc.title |
Binding of MetJ repressor to specific and nonspecific DNA and effect of S-adenosylmethionine
on these interactions.
|
|
dc.title.alternative |
|
|
dc.type |
Journal article |
|
duke.contributor.id |
Spicer, Leonard D|0114053 |
|
dc.description.version |
Version of Record |
|
duke.date.pubdate |
2010-4-20 |
|
duke.description.issue |
15 |
|
duke.description.volume |
49 |
|
dc.relation.journal |
Biochemistry |
|
pubs.author-url |
http://www.ncbi.nlm.nih.gov/pubmed/20196619 |
|
pubs.begin-page |
3289 |
|
pubs.end-page |
3295 |
|
pubs.issue |
15 |
|
pubs.organisational-group |
Basic Science Departments |
|
pubs.organisational-group |
Biochemistry |
|
pubs.organisational-group |
Clinical Science Departments |
|
pubs.organisational-group |
Duke |
|
pubs.organisational-group |
Duke Cancer Institute |
|
pubs.organisational-group |
Duke Human Vaccine Institute |
|
pubs.organisational-group |
Institutes and Centers |
|
pubs.organisational-group |
Radiology |
|
pubs.organisational-group |
School of Medicine |
|
pubs.publication-status |
Published |
|
pubs.volume |
49 |
|
dc.identifier.eissn |
1520-4995 |
|