title: Possible interaction between the bacterial transcription factor ArtA and the eukaryotic RNA polymerase III promoter.
abstract: Eukaryotic RNA polymerase III (RNAP III) transcribes tRNA genes and short interspersed elements that have internal promoters consisting of A- and B-blocks. The B-block binding subunit of the transcription initiation factor TFIIIC binds to the B-block. The mobile bacterial insertion sequence (IS) 1 contains a RNAP III promoter-like sequence, which stimulates bacterial transcription along with the bacterial ArtA protein. Here, the DNA-binding ability of ArtA was examined in vitro using a simple, newly developed method. Various DNA fragments, including RNAP III promoter fragments, were separately incubated with purified ArtA, and then loaded onto a polyacrylamide gel. Since DNAs bound by ArtA remain in the gel wells during electrophoresis, SDS was added into the wells at the electrophoresis halfway point. It was hypothesized that SDS would dissociate the DNA-ArtA complexes in the wells, and then the DNAs would begin to migrate. In fact, new bands appeared in all of the lanes at similar intensities, indicating that ArtA binds nonspecifically to DNA. Therefore, labeled wild-type RNAP III promoter fragments were incubated with either the unlabeled wild-type or mutant fragments and ArtA, and electrophoresed. The B-block(-like) sequences of IS1, a human Alu element, and an anuran tRNA gene were important for binding to ArtA. Additionally, in silico analyses revealed the presence of the RNAP III promoter-like structures in the IS1 isoforms and the IS3 family elements. These results suggest the presence of parts of the RNAP III transcription machinery in bacteria, and might imply that its prototype existed in the common ancestor.
authors.affiliation: Division of Microbiology, National Institute of Health Sciences, Tokyo, 158-8501, Japan. sachiko@nihs.go.jp.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Matsutani
mesh_terms: D020087:Alu Elements; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004251:DNA Transposable Elements; D056890:Eukaryota; D005786:Gene Expression Regulation; D006801:Humans; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012320:RNA Polymerase III / Q000235:genetics*; D012343:RNA, Transfer / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014981:Xenopus
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D014157:Transcription Factors; D012343:RNA, Transfer; D012320:RNA Polymerase III
keywords: Bacterial transposon; DNA–protein binding assay; Evolution; RNA polymerase III; Transcription machinery