DNA binding provides a molecular strap activating the adenovirus proteinase.
Human adenovirus proteinase (AVP) requires two cofactors for maximal activity: pVIc, a peptide derived from the C terminus of adenovirus precursor protein pVI, and the viral DNA. Synchrotron protein footprinting was used to map the solvent accessible cofactor binding sites and to identify conformational changes associated with the binding of cofactors to AVP. The binding of pVIc alone or pVIc and DNA together to AVP triggered significant conformational changes adjacent to the active site cleft sandwiched between the two AVP subdomains. In addition, upon binding of DNA to AVP, it was observed that specific residues on each of the two major subdomains were significantly protected from hydroxyl radicals. Based on the locations of these protected side-chain residues and conserved aromatic and positively charged residues within AVP, a three-dimensional model of DNA binding was constructed. The model indicated that DNA binding can alter the relative orientation of the two AVP domains leading to the partial activation of AVP by DNA. In addition, both pVIc and DNA may independently alter the active site conformation as well as drive it cooperatively to fully activate AVP.
Gupta
Sayan
S
Center for Synchrotron Biosciences, Department of Physiology & Biophysics, Albert Einstein College of Medicine, Bronx, NY 10461, USA
Mangel
Walter F
WF
McGrath
William J
WJ
Perek
Jennifer L
JL
Lee
Donna W
DW
Takamoto
Keiji
K
Chance
Mark R
MR
D000260:Adenoviruses, Human / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D024342:Amino Acids, Acidic; D024361:Amino Acids, Basic; D001665:Binding Sites; D004279:DNA, Viral / Q000378:metabolism*; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D023302:Protein Footprinting; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017356:Synchrotrons; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D024342:Amino Acids, Acidic; D024361:Amino Acids, Basic; D004279:DNA, Viral; D014764:Viral Proteins; D010450:Endopeptidases