Towards the Elucidation of the Protein Complexes Involved in Prokaryotic Origin Independent DNA Replication Restart-An ESI-MS Study

摘要

Origin independent replication restant in prokaryotes is accomplished by the primosome assembly proteins, PriA, PriB, PriC, DnaT, DnaB, DnaC and DnaG. Studies indicate two separate pathways of replication restart; one pathway initiated by PriA and the other by PriC. However, little is known about the order of assembly, stoichiometry, or structure of the primosome and even the individual functions of the proteins. We present the first attempt to use mass spectrometry to study the assembly of the proykaryotic replication restart proteins. Thus far, we have collected ESI-MS data for DnaT and PriB. Our data indicate that DuaT forms monomers and dimers. The data for PriB are consistent with the known crystal structure in that the protein forms a homodimer. Complex studies between the two proteins show interaction is mediated by the DNA.