The HPV16 E6 binding protein Tip-1 interacts with ARHGEF16, which activates Cdc42.

ABSTRACT:

Background

Guanidine exchange factor (GEF)-catalysed activation of Rho proteins such as Cdc42 has been shown to have a crucial role in cellular transformation, malignant progression and invasion. We have previously shown that the HPV16 E6 oncoprotein binds to the PDZ domain protein Tax-interacting-protein 1 (Tip-1) and we now report identification and functional analysis of a novel Tip-1 binding GEF.

Methods

Yeast two-hybrid, in vitro pull-down, site-directed mutagenesis, semiquantitative PCR, co-immunoprecipitation and western blotting were used to identify/confirm novel Tip-1 binding partners and analyse cellular expression levels. In vitro kinetic analyses of recombinant proteins, siRNA gene silencing and in cell assays were used to measure Rho protein activation.

Resu

SUBMITTER: Oliver AW 

PROVIDER: S-EPMC3031889 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature