Genetic analysis of the Φ174 DNA binding protein

Bryan Jennings, Bentley A Fane

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The X174 J protein is 37 amino acids in length and contains 12 basic residues. There are no acidic amino acids in the protein. The basic residues are concentrated in two clusters in the N-terminus which are separated by a proline-rich region. To investigate the morphogenetic functions of the J protein and possible mechanisms by which it may bind DNA, a genetic analysis was conducted. Lysine→leucine and arginine→leucine substitutions were generated within the basic amino acid clusters. At least three substitutions were required to eliminate viability in vivo. Lethal mutants with three or four substitutions exhibit dominant lethal phenotypes, indicating that the mutant proteins retain enough function to interfere with productive assembly. In cells infected with a dominant lethal mutant, noninfectious packaged particles were produced. Infectivity can be restored by second-site suppressors in the viral coat protein which disrupt polar interactions atop the threefold axis of symmetry in the capsid. The viability of strains containing compensating frameshift mutations within the proline-rich region suggests that only the proline residues in this segment are critical for efficient function.

Original languageEnglish (US)
Pages (from-to)370-377
Number of pages8
JournalVirology
Volume227
Issue number2
DOIs
StatePublished - Jan 20 1997
Externally publishedYes

Fingerprint

DNA-Binding Proteins
Proline
Acidic Amino Acids
Basic Amino Acids
Frameshift Mutation
Proteins
Capsid
Capsid Proteins
Mutant Proteins
Phenotype
Amino Acids
DNA

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Genetic analysis of the Φ174 DNA binding protein. / Jennings, Bryan; Fane, Bentley A.

In: Virology, Vol. 227, No. 2, 20.01.1997, p. 370-377.

Research output: Contribution to journalArticle

Jennings, Bryan ; Fane, Bentley A. / Genetic analysis of the Φ174 DNA binding protein. In: Virology. 1997 ; Vol. 227, No. 2. pp. 370-377.
@article{5a405b8e923049798ca6eb20a57c28e0,
title = "Genetic analysis of the Φ174 DNA binding protein",
abstract = "The X174 J protein is 37 amino acids in length and contains 12 basic residues. There are no acidic amino acids in the protein. The basic residues are concentrated in two clusters in the N-terminus which are separated by a proline-rich region. To investigate the morphogenetic functions of the J protein and possible mechanisms by which it may bind DNA, a genetic analysis was conducted. Lysine→leucine and arginine→leucine substitutions were generated within the basic amino acid clusters. At least three substitutions were required to eliminate viability in vivo. Lethal mutants with three or four substitutions exhibit dominant lethal phenotypes, indicating that the mutant proteins retain enough function to interfere with productive assembly. In cells infected with a dominant lethal mutant, noninfectious packaged particles were produced. Infectivity can be restored by second-site suppressors in the viral coat protein which disrupt polar interactions atop the threefold axis of symmetry in the capsid. The viability of strains containing compensating frameshift mutations within the proline-rich region suggests that only the proline residues in this segment are critical for efficient function.",
author = "Bryan Jennings and Fane, {Bentley A}",
year = "1997",
month = "1",
day = "20",
doi = "10.1006/viro.1996.8351",
language = "English (US)",
volume = "227",
pages = "370--377",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Genetic analysis of the Φ174 DNA binding protein

AU - Jennings, Bryan

AU - Fane, Bentley A

PY - 1997/1/20

Y1 - 1997/1/20

N2 - The X174 J protein is 37 amino acids in length and contains 12 basic residues. There are no acidic amino acids in the protein. The basic residues are concentrated in two clusters in the N-terminus which are separated by a proline-rich region. To investigate the morphogenetic functions of the J protein and possible mechanisms by which it may bind DNA, a genetic analysis was conducted. Lysine→leucine and arginine→leucine substitutions were generated within the basic amino acid clusters. At least three substitutions were required to eliminate viability in vivo. Lethal mutants with three or four substitutions exhibit dominant lethal phenotypes, indicating that the mutant proteins retain enough function to interfere with productive assembly. In cells infected with a dominant lethal mutant, noninfectious packaged particles were produced. Infectivity can be restored by second-site suppressors in the viral coat protein which disrupt polar interactions atop the threefold axis of symmetry in the capsid. The viability of strains containing compensating frameshift mutations within the proline-rich region suggests that only the proline residues in this segment are critical for efficient function.

AB - The X174 J protein is 37 amino acids in length and contains 12 basic residues. There are no acidic amino acids in the protein. The basic residues are concentrated in two clusters in the N-terminus which are separated by a proline-rich region. To investigate the morphogenetic functions of the J protein and possible mechanisms by which it may bind DNA, a genetic analysis was conducted. Lysine→leucine and arginine→leucine substitutions were generated within the basic amino acid clusters. At least three substitutions were required to eliminate viability in vivo. Lethal mutants with three or four substitutions exhibit dominant lethal phenotypes, indicating that the mutant proteins retain enough function to interfere with productive assembly. In cells infected with a dominant lethal mutant, noninfectious packaged particles were produced. Infectivity can be restored by second-site suppressors in the viral coat protein which disrupt polar interactions atop the threefold axis of symmetry in the capsid. The viability of strains containing compensating frameshift mutations within the proline-rich region suggests that only the proline residues in this segment are critical for efficient function.

UR - http://www.scopus.com/inward/record.url?scp=0031579233&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031579233&partnerID=8YFLogxK

U2 - 10.1006/viro.1996.8351

DO - 10.1006/viro.1996.8351

M3 - Article

VL - 227

SP - 370

EP - 377

JO - Virology

JF - Virology

SN - 0042-6822

IS - 2

ER -