The complexes were competed selleck chemicals llc out with the corresponding unlabeled UP1B or UP2 consensus GATA oligonucleotides, but not with mutant versions of these sequences. In addition, we observed supershift of the UP1B complex in the presence of GATA4 and GATA6 antibodies (Figure 6C), consistent with the presence of GATA4/6 in the complex. Supershift was also noted with the UP2 complex in the presence of GATA4 antibody, but not with IgG control; however the presence of GATA6 antibody resulted in substantial immunodepletion of the UP2 complex (Figu
Human adenoviruses (Ads) cause a wide range of diseases [1-3] but it is incompletely known how virus structure relates to infection. Ad particles consist of an icosahedral capsid enclosing a linear-double stranded DNA genome.

The outer capsid is made of hexon (protein II), the penton base at the vertices (protein III), the protruding trimeric fibers (protein IV), and various minor proteins, IIIa, VI, VIII and IX. The inner core contains the double-stranded DNA with condensing proteins VII, V, and X, two copies of the terminal protein at the 5′ ends of the DNA, the IVa2 core protein, and about 10 copies of the 23 kDa protease L3/p23. L3/p23 is highly conserved across human Ads, and has important roles in virion morphogenesis and entry [4]. It cleaves substrates at glycine and isoleucine-containing consensus sites [5,6], and requires cofactors for optimal activity [7-9]. During virion assembly, L3/p23 cleaves six structural precursor (p) proteins, pIIIa, pVI, pVII, pVIII, pX, the preterminal protein (pTP), and possibly the L1-52/55K scaffolding protein [10].

L3/p23 cleaves V and pVII at putative cleavage sites, and hexon and pVI at degenerate cleavage sites in vitro [11]. The isolation of the temperature-sensitive (ts) Ad2-ts1 suggested that L3/p23 encoded a protease [12]. Ad2-ts1 is defective in protease packaging, and virion processing at the nonpermissive temperature (40��C) [13]. The mutation was mapped to P137L of L3/p23 [14], and eliminated by spontaneous reversions of the C22187T mutation in L3/p23 [15]. Yet, the recombinant P137L protease is catalytically active [16,17], and the ts1-phenotype rescued by adding a protease-activating peptide of the C-terminus of pVI to infected cells [13]. It is unknown if secondary mutations in Ad2-ts1 act synergistically Entinostat with P137L, and contribute to the phenotype. To dissect the complex Ad2-ts1 entry phenotype [18,19], we sequenced the structural proteins, the packaging-related proteins and the origins of replication of Ad2-ts1.