Interestingly, p53 activation induces caspase-6 which is responsible for caspase-mediated HIPK2 cleavage at positions 916 and 977 [19]. This C-terminus truncated HIPK2 results in a hyperactive kinase which potentiates p53Ser46 phosphorylation and activation of apoptosis learn more and eventually is degraded. Thus, caspase-resistant HIPK2 mutants induce apoptosis less efficiently than wild-type [19]. These findings suggest a tight regulation of HIPK2 in a p53-dependent manner, a regulatory loop similar to the elimination of ERK2 kinase by

a p53-induced apoptotic program, in order to prevent ERK-mediated cell proliferation in the presence of activated p53 [20]. HIPK2 is a critical activator of p53 function in response to drugs as substantiate by experiments of HIPK2 gene silencing by small interference RNA (siRNA). HIPK2 knockdown impairs p53 pro-apoptotic gene transcription in response to drugs and predisposes to chemoresistance [14] and increased tumor growth in vivo[21]. HIPK2 knockdown contributes to p53 inactivation by different means other than by direct impairment of p53Ser46 phosphorylation. cDNA microarray BYL719 molecular weight of colon cancer cells with chronic depletion of HIPK2 function by siRNA [22], showed upregulation of two novel targets of HIPK2 corepressor function that are involved in p53 deregulation, that is, Nox1 and

MT2A. Thus, HIPK2 has been shown to repress Nox1 promoter activity [23]. Nox1 is a homolog of the catalytic subunit of the superoxide-generating NADPH-oxidase that is often

overexpressed in tumors and is involved in tumor progression and angiogenesis [24]. HIPK2 knockdown induces Nox1 upregulation and Nox1 overexpression impairs p53 apoptotic transcriptional activity by inducing p53Lys382 deacetylation [23]. Interestingly, chronic HIPK2 depletion leads to p53 protein misfolding, as assessed by immunoprecipitation studies with conformation-specific p53 antibodies, that impairs p53/DNA binding and p53 transcriptional activity [22]. This p53 misfolding, in colon and breast cancer cells, could be, at least in part, Selleckchem Luminespib ascribed to metallothionein 2A (MT2A) upregulation upon HIPK2 depletion [25]. Thus, MT2A depletion by siRNA, restores wtp53 native conformation TCL and p53 function in response to drugs, in HIPK2 knockdown cells [25]. Metallothionein is a family of at least 10 conserved isoforms of metal-binding cysteine-rich proteins with a potential role in homeostasis of essential metals [26]. MTs upregulation has been found in several human tumors including breast, colon, liver, and lung, and supports a role for MTs in acquired drug resistance [27]. In most cell types, zinc is often sequestered through binding to MTs, keeping free zinc concentrations fairly low that could account for lack of function in a typical zinc-sensitive protein, such as p53 [28].