Hybridization selection of covalent nucleic acid-protein complexes. 2. Cross-linking of proteins to specific Escherichia coli mRNAs and DNA sequences by formaldehyde treatment of intact cells.
Proteins cross-linked to pBR322 mRNAs and DNA by formaldehyde treatment of intact Escherichia coli cells have been detected with the use of a novel detection method. Among the proteins cross-linked to pBR322 mRNAs were S1, S21, and at least six other proteins of the small ribosomal subunit, initiation factor 1, elongation factor (EF) Tu, and very small amounts of EF-G and EF-Ts. The single strand binding protein, the HU-proteins, and RNA polymerase subunits alpha and beta were among the proteins cross-linked to pBR322 DNA. The results obtained suggest that the procedures described, can also be used to study interactions between different nucleic acid-bound polypeptides. The results are discussed in relation to the working mechanism of formaldehyde, and are compared to the results obtained with cross-linking induced by ultraviolet light. The methods presented should also be of use for the study of nucleic acid-protein interactions in other organisms.
D001426:Bacterial Proteins / Q000378:metabolism; D002846:Chromatography, Affinity; D004269:DNA, Bacterial / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D015977:Eukaryotic Initiation Factor-1; D005557:Formaldehyde / Q000494:pharmacology*; D008970:Molecular Weight; D020653:Peptide Elongation Factor G; D010444:Peptide Elongation Factor Tu; D010445:Peptide Elongation Factors / Q000378:metabolism; D039861:Prokaryotic Initiation Factor-1; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D015977:Eukaryotic Initiation Factor-1; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; D039861:Prokaryotic Initiation Factor-1; D011506:Proteins; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D005557:Formaldehyde; D012321:DNA-Directed RNA Polymerases; D010444:Peptide Elongation Factor Tu