Nucleic acid-binding regions of the second-largest subunit of Drosophila RNA polymerase II identified by Southwestern blotting

• Published in: FEBS letters Vol. 344; no. 2; pp. 166 - 170
• Main Authors: Kontermann, Roland E., Bautz, Ekkehard K.F.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 16.05.1994; Elsevier
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Base Sequence; beta-Galactosidase - genetics; Binding Sites; Biological and medical sciences; Blotting, Southern; Blotting, Western; DNA - metabolism; Drosophila melanogaster; Drosophila melanogaster - enzymology; Enzymes and enzyme inhibitors; Escherichia coli; Fundamental and applied biological sciences. Psychology; Fusion protein; Isoelectric Point; Molecular Sequence Data; Non-specific DNA binding; Recombinant Fusion Proteins - metabolism; RNA polymerase; RNA Polymerase II - chemistry; RNA Polymerase II - genetics; RNA Polymerase II - metabolism; Sequence Alignment; Southwestern blotting; Transcription; Transferases; Non-specific DNA binding; RNA polymerase; Transcription; Fusion protein; Southwestern blotting; Enzyme; Insecta; Transferases; Binding site; Subunit; DNA-directed RNA polymerase; Drosophilidae; Nucleotidyltransferases; Arthropoda; DNA; Invertebrata; Localization; Drosophila melanogaster; Diptera
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(94)00380-7

Analysing overlapping bacterially expressed fragments of the second-largest subunit of Drosophila melanogaster RNA polymerase II in Southwestern DNA binding assays we have identified regions that have the potential to bind nucleic acids non-specifically. A region exhibiting strong DNA binding is located in the N-terminal part of the molecule (amino acids 357–504) and some weak DNA binding is observed for the C-terminal part (amino acids 860–1160). The non-specific DNA binding behavior of these regions is similar to that of the native enzyme. Most of the known mutations responsible for rifampicin resistance map to a region of the Escherichia coli β subunit corresponding to the N-terminal nucleic acid-binding region, indirectly supporting the notion that this region participates in interaction with the RNA transcript in ternary complexes.