title: Conversion of the E1A Cys4 zinc finger to a nonfunctional His2,Cys2 zinc finger by a single point mutation.
abstract: Trans-activation by the adenovirus E1A 289R protein requires a zinc finger defined by Cys-154, Cys-157, Cys-171, and Cys-174. Whereas individually replacing the four cysteine residues with serines resulted in a loss of transactivation, only three of the Cys----Ser mutants (C157S, C171S, and C174S) lost the ability to bind Zn(II). X-ray absorption fine structure analysis revealed that, in the wild-type protein, Zn(II) is coordinated by four cysteine residues whereas in the C154S mutant, Zn(II) is coordinated by two histidines and two cysteines. The mutant protein probably retains, as ligands, two cysteines on the right side of the zinc finger (Cys-171 and Cys-174) and recruits two of the four histidines on the left side (His-149, His-152, His-158, and His-160), despite the presence of Cys-157. This finding may shed light on the general structural requirements of zinc fingers.
authors.affiliation: Institute for Structural Functional Studies, Philadelphia, PA 19104.
authors.forename: L C
authors.initials: LC
authors.lastname: Webster
authors.forename: K
authors.initials: K
authors.lastname: Zhang
authors.forename: B
authors.initials: B
authors.lastname: Chance
authors.forename: I
authors.initials: I
authors.lastname: Ayene
authors.forename: J S
authors.initials: JS
authors.lastname: Culp
authors.forename: W J
authors.initials: WJ
authors.lastname: Huang
authors.forename: F Y
authors.initials: FY
authors.lastname: Wu
authors.forename: R P
authors.initials: RP
authors.lastname: Ricciardi
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D017456:Adenovirus Early Proteins; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D003545:Cysteine; D004268:DNA-Binding Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010957:Plasmids; D011487:Protein Conformation; D015533:Transcriptional Activation; D014162:Transfection; D015032:Zinc / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D017456:Adenovirus Early Proteins; D003062:Codon; D004268:DNA-Binding Proteins; D009856:Oncogene Proteins, Viral; D015032:Zinc; D003545:Cysteine