TY - JOUR
T1 - Binding of the σ70 protein to the core subunits of Escherichia coli RNA polymerase, studied by iron-EDTA protein footprinting
AU - Greiner, Douglas P.
AU - Hughes, Karin A.
AU - Gunasekera, Angelo H.
AU - Meares, Claude F.
PY - 1996/1/9
Y1 - 1996/1/9
N2 - We have used a nonspecific protein cleaving reagent to map the interactions between subunits of the multisubunit enzyme RNA polymerase (Escherichia coli). We developed suitable conditions for using an untethered Fe-EDTA reagent, which does not bind significantly to proteins. Comparison of the cleaved fragments of the subunits from the core enzyme (α2ββ′) and the holoenzyme (core + σ70) shows that absence of the σ70 subunit is associated with the appearance of several cleavage sites on the subunits β (within 10 residues of sequence positions 745, 764, 795, and 812) and β′ (within 10 residues of sequence positions 581, 613, and 728). A cleavage site near β residue 604 is present in the holoenzyme but absent in the core, demonstrating that a conformational change occurs when σ70 binds. No differences are observed for the α subunit.
AB - We have used a nonspecific protein cleaving reagent to map the interactions between subunits of the multisubunit enzyme RNA polymerase (Escherichia coli). We developed suitable conditions for using an untethered Fe-EDTA reagent, which does not bind significantly to proteins. Comparison of the cleaved fragments of the subunits from the core enzyme (α2ββ′) and the holoenzyme (core + σ70) shows that absence of the σ70 subunit is associated with the appearance of several cleavage sites on the subunits β (within 10 residues of sequence positions 745, 764, 795, and 812) and β′ (within 10 residues of sequence positions 581, 613, and 728). A cleavage site near β residue 604 is present in the holoenzyme but absent in the core, demonstrating that a conformational change occurs when σ70 binds. No differences are observed for the α subunit.
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M3 - Article
C2 - 8552678
AN - SCOPUS:0029664584
VL - 93
SP - 71
EP - 75
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 1
ER -