In vitro binding of the bacteriophage f1 gene V protein to the gene II RNA-operator and its DNA analog

• Published in: Nucleic acids research 1989-09, Vol.17 (18), p.7333-7344
• Main Authors: MICHEL, B, ZINDER, N. D
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Coliphages - genetics; DNA Mutational Analysis; Dna, deoxyribonucleoproteins; DNA, Single-Stranded - metabolism; DNA-Binding Proteins - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Viral; In Vitro Techniques; Nucleic acids; Oligonucleotides - metabolism; Operator Regions, Genetic; Protein Biosynthesis; Repressor Proteins - metabolism; RNA, Messenger - metabolism; RNA, Viral - metabolism; Transcription Factors - metabolism; Viral Proteins - genetics
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/17.18.7333

We have investigated the binding of the f1 single-stranded DNA-binding protein (gene V protein) to DNA oligonucleotides and RNA synthesized in vitro. The first 16 nucleotides of the f1 gene II mRNA leader sequence were previously identified as the gene II RNA-operator; the target to which the gene V protein binds to repress gene II translation. Using a gel retardation assay, we find that the preferential binding of gene V protein to an RNA carrying the gene II RNA-operator sequence is affected by mutations which abolish gene II translational repression in vivo. In vitro, gene V protein also binds preferentially to a DNA oligonucleotide whose sequence is the DNA analog of the wild-type gene II RNA-operator. Therefore, the gene V protein recognizes the gene II mRNA operator sequence when present in either an RNA or DNA context.