Evidence for the complex formation between reverse transcriptase and multicopy single-stranded DNA in retron EC83.
Multicopy single-stranded DNA (msDNA) is produced by reverse transcriptase (RT) and is mutagenic in E. coli. In order to study the interaction between msDNA and RT, an RT open reading frame of retron EC83 was fused with maltose binding protein. The fusion protein, MBP-RT, was active in msDNA synthesis. The msDNA was copurified with MBP-RT by amylose affinity column. When purified msDNA was mixed with MBP-RT, the msDNA formed a complex with the fusion protein. These results show that msDNA is present as a complex with RT in vivo and it is able to form such a complex in vitro. We suggest that RT may compete with MutS protein for binding to msDNA and thereby controls mutagenesis by msDNA.
Jeong
D W
DW
Department of Microbiology, College of Natural Sciences, Gyeongsang National University, Chinju, Korea.
Kim
K
K
Lim
D
D
D018528:ATP-Binding Cassette Transporters; D000251:Adenosine Triphosphatases; D001426:Bacterial Proteins / Q000378:metabolism; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D008320:Maltose / Q000378:metabolism; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D051719:MutS DNA Mismatch-Binding Protein; D016366:Open Reading Frames; D011485:Protein Binding; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D018626:Retroelements / Q000235:genetics*
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D012329:RNA, Bacterial; D011993:Recombinant Fusion Proteins; D018626:Retroelements; C461599:maltose transport system, E coli; C058681:msDNA; D008320:Maltose; D012194:RNA-Directed DNA Polymerase; D000251:Adenosine Triphosphatases; D051719:MutS DNA Mismatch-Binding Protein; C497175:MutS protein, E coli