Single-stranded DNA binding proteins (SSBs) from prokaryotic transmissible plasmids.
The DNA and protein sequences of single-stranded DNA binding proteins (SSBs) encoded by the plP71a, plP231a, and R64 conjugative plasmids have been determined and compared to Escherichia coli SSB and the SSB encoded by F-plasmid. Although the amino acid sequences of all of these proteins are highly conserved within the NH2-terminal two-thirds of the protein, they diverge in the COOH-terminal third region. A number of amino acid residues which have previously been implicated as being either directly or indirectly involved in DNA binding are conserved in all of these SSBs. These residues include Trp-40, Trp-54, Trp-88, His-55, and Phe-60. On the basis of these sequence comparisons and DNA binding studies, a role for Tyr-70 in DNA binding is suggested for the first time. Although the COOH-terminal third of these proteins diverges more than their NH2-terminal regions, the COOH-terminal five amino acid residues of all five of these proteins are identical. In addition, all of these proteins share the characteristic property of having a protease resistant, NH2-terminal core and an acidic COOH-terminal region. Despite the high degree of sequence homology among the plasmid SSB proteins, the F-plasmid SSB appears unique in that it was the only SSB tested that neither bound well to poly(dA) nor was able to stimulate DNA polymerase III holoenzyme elongation rates. Poly [d(A-T)] melting studies suggest that at least three of the plasmid encoded SSBs are better helix-destabilizing proteins than is the E. coli SSB protein.
Ruvolo
P P
PP
Department of Molecular Biology and Genetics, Albert Einstein College of Medicine, Bronx, New York 10461.
Keating
K M
KM
Williams
K R
KR
Chase
J W
JW
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003227:Conjugation, Genetic; D004258:DNA Polymerase III / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011061:Poly A / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013696:Temperature
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D011061:Poly A; C015465:poly(dA); D004258:DNA Polymerase III