J. Li et Ss. Broyles, THE DNA-BINDING DOMAIN OF THE VACCINIA VIRUS EARLY TRANSCRIPTION FACTOR SMALL-SUBUNIT IS AN EXTENDED HELICASE-LIKE MOTIF, Nucleic acids research, 23(9), 1995, pp. 1590-1596
The vaccinia virus early transcription factor (VETF) is an ATP-depende
nt activator of the early class of viral genes, VETF is a heterodimeri
c protein that binds an initiator-like element surrounding the start s
ite of transcription. Previous studies indicated that the small subuni
t of VETF contacts the promoter DNA, We have taken a mutational approa
ch to determine sequences in the VETF small subunit that are important
for DNA binding, Two types of sequences were targeted for mutation: o
nes resembling motifs that are conserved in the nucleic acid helicase
family and positively charged residues in predicted alpha-helices, Mut
ations affecting transcription activation were clustered in two region
s, One mutation that impaired DNA binding is located near the N-termin
us within the putative ATP-binding pocket that comprises helicase doma
in I. DNA binding was also severely reduced by mutations in a sequence
resembling helicase domain VI and two putative alpha-helices that fla
nk this domain in the C-terminal third of the polypeptide, These resul
ts indicate that the DNA binding domain in the small subunit of VETF i
s not isolated within a separable domain as is the case with most tran
scription factors, but rather, spans most of the length of the 637 res
idue polypeptide. A model for VETF structure is suggested in which the
active site for ATP hydrolysis is integrated within an extended DNA-b
inding domain such that the structure and function of each domain infl
uences that of the other.