title: Domain effects on the DNA-interactive properties of bacteriophage T4 gene 32 protein.
abstract: Bacteriophage T4 gene 32 protein, a model for single-strand specific nucleic acid-binding proteins, consists of three structurally and functionally distinct domains. We have studied the effects of the N and C domains on the protein structure and its nucleic acid-interactive properties. Although the presence of the C domain decreases the proteolytic susceptibility of the core (central) domain, quenching of the core tryptophan fluorescence by iodide is unaltered by the presence of the terminal domains. These results suggest that the overall conformation of the core domain remains largely independent of the flanking domains. Removal of the N or the C terminus does not abolish the DNA renaturation activity of the protein. However, intact protein and its three truncated forms differ in DNA helix-destabilizing activity. The C domain alone is responsible for the kinetic barrier to natural DNA helix destabilization seen with intact protein. Intact protein and core domain potentiate the DNA helix-destabilizing activity of truncated protein lacking only the C domain (*I), enhancing the observed hyperchromicity while increasing the melting temperature. Proteolysis experiments suggest that the affinity of core domain for single-stranded DNA is increased in the presence of *I. We propose that *I can "mingle" with intact protein or core domain while bound to single-stranded DNA.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, Baltimore County, Baltimore 21250, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Waidner
authors.forename: E K
authors.initials: EK
authors.lastname: Flynn
authors.forename: M
authors.initials: M
authors.lastname: Wu
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.forename: R L
authors.initials: RL
authors.lastname: Karpel
mesh_terms: D017122:Bacteriophage T4; D018736:DNA Adducts; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D012697:Serine Endopeptidases; D014364:Tryptophan; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D018736:DNA Adducts; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014764:Viral Proteins; C060035:gp32 protein, Enterobacteria phage T4; D014364:Tryptophan; C044499:submandibular proteinase A; D012697:Serine Endopeptidases