Abstract
Gene V protein of bacteriophage Ff (M13, f1, fd) is a master regulator of phage DNA replication and phage mRNA translation. It exerts these two functions by binding to single-stranded viral DNA or to specific sequences in the 5' ends of its target mRNAs, respectively. To study the structure/function relationship of gene V protein, M13 gene V was inserted in a phagemid expression vector and a library of missense and nonsense mutants was constructed by random chemical mutagenesis. Phagemids encoding gene V proteins with decreased biological activities were selected at the nucleotide sequences of their gene V fragments were determined. Furthermore, the mutant proteins were characterized both with respect to their ability to inhibit the production of phagemid DNA transducing particles and their ability to repress the translation of a chimeric lacZ receptor gene whose expression is controlled by the promoter and translational initiation signals of M13 gene II. From the data obtained, it can be deduced that the mechanism by which gene V protein binds to single-stranded DNA differs from the mechanism by which it binds to its target sequences in the gene II mRNA.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1003-1014 |
| Number of pages | 12 |
| Journal | European Journal of Biochemistry |
| Volume | 204 |
| Issue number | 3 |
| State | Published - 1992 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
Cite this
Selection and characterization of randomly produced mutants of gene V protein of bacteriophage M13. / Stassen, A. P M; Zaman, G. J R; Van Deursen, Jan; Schoenmakers; Konings, R. N H.
In: European Journal of Biochemistry, Vol. 204, No. 3, 1992, p. 1003-1014.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Selection and characterization of randomly produced mutants of gene V protein of bacteriophage M13
AU - Stassen, A. P M
AU - Zaman, G. J R
AU - Van Deursen, Jan
AU - Schoenmakers,
AU - Konings, R. N H
PY - 1992
Y1 - 1992
N2 - Gene V protein of bacteriophage Ff (M13, f1, fd) is a master regulator of phage DNA replication and phage mRNA translation. It exerts these two functions by binding to single-stranded viral DNA or to specific sequences in the 5' ends of its target mRNAs, respectively. To study the structure/function relationship of gene V protein, M13 gene V was inserted in a phagemid expression vector and a library of missense and nonsense mutants was constructed by random chemical mutagenesis. Phagemids encoding gene V proteins with decreased biological activities were selected at the nucleotide sequences of their gene V fragments were determined. Furthermore, the mutant proteins were characterized both with respect to their ability to inhibit the production of phagemid DNA transducing particles and their ability to repress the translation of a chimeric lacZ receptor gene whose expression is controlled by the promoter and translational initiation signals of M13 gene II. From the data obtained, it can be deduced that the mechanism by which gene V protein binds to single-stranded DNA differs from the mechanism by which it binds to its target sequences in the gene II mRNA.
AB - Gene V protein of bacteriophage Ff (M13, f1, fd) is a master regulator of phage DNA replication and phage mRNA translation. It exerts these two functions by binding to single-stranded viral DNA or to specific sequences in the 5' ends of its target mRNAs, respectively. To study the structure/function relationship of gene V protein, M13 gene V was inserted in a phagemid expression vector and a library of missense and nonsense mutants was constructed by random chemical mutagenesis. Phagemids encoding gene V proteins with decreased biological activities were selected at the nucleotide sequences of their gene V fragments were determined. Furthermore, the mutant proteins were characterized both with respect to their ability to inhibit the production of phagemid DNA transducing particles and their ability to repress the translation of a chimeric lacZ receptor gene whose expression is controlled by the promoter and translational initiation signals of M13 gene II. From the data obtained, it can be deduced that the mechanism by which gene V protein binds to single-stranded DNA differs from the mechanism by which it binds to its target sequences in the gene II mRNA.
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M3 - Article
C2 - 1551382
AN - SCOPUS:0026607453
VL - 204
SP - 1003
EP - 1014
JO - FEBS Journal
JF - FEBS Journal
SN - 1742-464X
IS - 3
ER -