[Visualization of bacteriophage T7 RNA-polymerase complexes with DNA template in the process of transcription elongation].
Complexes of bacteriophage T7 RNA polymerase (T7 RNAP) with a DNA template (which contains a promoter and internal terminator of T7 RNAP) in transcription elongation were visualized by atomic force microscopy (AFM). Images of specific stable complexes of T7 RNAP with a terminal fragment of DNA template located at a distance of 200 bp from the T7 RNAP promoter were obtained for single molecules. Complexes of a single DNA template molecule with 2-3 T7 RNAP molecules corresponding to stages of initiation, elongation and termination of transcription were visualized under the elimination of nonspecific DNA-protein binding. Detailes of specific and nonspecific complex formation for the T7 RNAP--DNA system during initiation and transcription elongation are discussed.
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D017123:Bacteriophage T7 / Q000201:enzymology / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism* / Q000648:ultrastructure; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism* / Q000648:ultrastructure; D004587:Electrophoresis, Agar Gel; D018625:Microscopy, Atomic Force; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D035602:Transcriptional Elongation Factors / Q000378:metabolism* / Q000648:ultrastructure; D014764:Viral Proteins / Q000378:metabolism* / Q000648:ultrastructure
D035602:Transcriptional Elongation Factors; D014764:Viral Proteins; D004247:DNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases