The minute virus of mice capsid specifically recognizes the 3' hairpin structure of the viral replicative-form DNA: mapping of the binding site by hydroxyl radical footprinting.

ABSTRACT Parvoviral rolling hairpin replication generates palindromic genomic concatemers whose junctions are resolved to give unit-length genomes by a process involving DNA replication initiated at origins derived from each viral telomere. The left-end origin of minute virus of mice (MVM), oriL, contains binding sites for the viral initiator nickase, NS1, and parvovirus initiation factor (PIF), a member of the emerging KDWK family of transcription factors. oriL is generated as an active form, oriLTC, and as an inactive form, oriLGAA, which contains a single additional nucleotide inserted between the NS1 and PIF sites. Here we examined the interactions on oriLTC which lead to activation of NS1 by PIF. The two subunits of PIF, p79 and p96, cooperatively bind two ACGT half-sites, which can be flexibly spaced. When coexpressed from recombinant baculoviruses, the PIF subunits preferentially form heterodimers which, in the presence of ATP, show cooperative binding with NS1 on oriL, but this interaction is preferentially enhanced on oriLTC compared to oriLGAA. Without ATP, NS1 is unable to bind stably to its cognate site, but PIF facilitates this interaction, rendering the NS1 binding site, but not the nick site, resistant to DNase I. Varying the spacing of the PIF half-sites shows that the distance between the NS1 binding site and the NS1-proximal half-site is critical for nickase activation, whereas the position of the distal half-site is unimportant. When expressed separately, both PIF subunits form homodimers that bind site specifically to oriL, but only complexes containing p79 activate the NS1 nickase function.

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NF-Y controls transcription of the minute virus of mice P4 promoter through interaction with an unusual binding site.

Journal of Virology ◽

10.1128/jvi.69.1.239-246.1995 ◽

1995 ◽

Vol 69 (1) ◽

pp. 239-246 ◽

Cited By ~ 14

Author(s):

Z Gu ◽

S Plaza ◽

M Perros ◽

C Cziepluch ◽

J Rommelaere ◽

...

Keyword(s):

Binding Site ◽

Minute Virus Of Mice ◽

Minute Virus

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The BldD Protein from Streptomyces coelicolor Is a DNA-Binding Protein

Journal of Bacteriology ◽

10.1128/jb.181.21.6832-6835.1999 ◽

1999 ◽

Vol 181 (21) ◽

pp. 6832-6835 ◽

Cited By ~ 34

Author(s):

Marie A. Elliot ◽

Brenda K. Leskiw

Keyword(s):

Escherichia Coli ◽

Hydroxyl Radical ◽

Binding Site ◽

Promoter Region ◽

Streptomyces Coelicolor ◽

Dna Binding Protein ◽

Mobility Shift ◽

Gel Mobility Shift ◽

A Site ◽

Hydroxyl Radical Footprinting

ABSTRACT Gel mobility shift assays with His-tagged BldD isolated fromEscherichia coli have illustrated that BldD is capable of specifically recognizing its own promoter region. DNase I and hydroxyl radical footprinting assays have served to delimit the BldD binding site, revealing that BldD recognizes and binds to a site just upstream from, and overlapping with, the −10 region of the promoter. How BldD binds to its promoter and the effect this binding has on the expression of BldD are discussed.

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