10 nm RecA protein filaments formed in the presence of Mg2+ and ATP gamma S may contain RNA.
Filaments formed by the polymerization of RecA protein along DNA in the presence of Mg2+ and adenosine 5'-0-(3-thiotriphosphate) (ATP gamma S) are seen by electron microscopy to have a 10 nm diameter with a 9 nm helical repeat. When certain preparations of apparently pure RecA protein are incubated with Mg2+ and ATP gamma S in the absence of nucleic acid for extended times, very long filaments with the same 10 nm diameter and 9 nm axial repeat are seen. We show here that these long 10 nm filaments can contain RNA which is present as a contaminant of the RecA protein and poly(A) which is synthesized during the incubations by an activity that is apparently polynucleotide phosphorylase. RecA protein purified by a procedure developed in this laboratory did not contain RNA and did not form these very long 10 nm filaments. However, when exogenous RNA was added to this protein, 10 nm filament formation was observed.
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D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology*; D008854:Microscopy, Electron; D011485:Protein Binding; D012329:RNA, Bacterial / Q000378:metabolism*; D011938:Rec A Recombinases / Q000378:metabolism*; D013873:Thionucleotides / Q000494:pharmacology*
D046911:Macromolecular Substances; D012329:RNA, Bacterial; D013873:Thionucleotides; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D011938:Rec A Recombinases; D008274:Magnesium