Myxococcus xanthus msDNA.Mx162 exists as a complex with proteins.
Myxococcus xanthus, a myxobacterium, contains a peculiar branched RNA-linked DNA called msDNA. Reverse transcriptase has been shown to be required for the production of msDNA. Existence of proteins that bind to one of the two msDNAs in M. xanthus, msDNA.Mx162, was examined by gel retardation assays. Total cell-free extract yielded two distinct retarded bands. Both bands were sensitive to treatment with proteinase K, indicating that there is a protein(s) that is able to bind to msDNA. Further, the formation of the bands was inhibited by the addition of nonradioactive msDNA but not by a large excess of poly(dA) in the presence of a 5000-fold excess of poly(dI.dC).poly(dI.dC). In vivo footprinting using dimethyl sulfate revealed that the deoxynucleotide stretch from 60 to 161 is protected. When a M. xanthus cell lysate was centrifuged in a 16-30% glycerol gradient, msDNA was found to sediment in two peaks: a major peak corresponding in size to 14 S, and a minor one at 5 S. These results indicate that msDNA.Mx162 exists as a complex with specific proteins in the cell.
Viswanathan
M
M
Department of Biochemistry, Robert Wood Johnson Medical School at Rutgers, University of Medicine and Dentistry of New Jersey, Piscataway 08854.
Inouye
M
M
Inouye
S
S
D001483:Base Sequence; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009231:Myxococcales / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D009710:Nucleotide Mapping; D011485:Protein Binding