Anti-VRK3 (full length) polyclonal antibody

Vaccinia-related kinase 3 (VRK3), previously characterized as a direct activator of vaccinia HI-related (VHR) phosphatase, inactivates extracellular signal-regulated kinase (ERK) in the nucleus of neuronal cells. Here we show that VRK3 is expressed in various other rodent tissues and in embryos, and regulates VHR phosphatase activity in these tissues. We observed colocalization of VRK3 and VHR in the testis tissue and could detect protein complex containing VRK3, VHR and ERK in immunoprecipitation analysis. Notably, the addition of recombinant VRK3 protein to total protein lysates, obtained either from adult tissues or embryos, enhanced the phosphatase activity of VHR, but not the activity of MKP3. The results further indicate that the VHR-VRK3 complex is a phosphatase-active form. In addition, we found that VRK3 can regulate EGF-induced cellular growth signaling that is mediated by ERK activation. Our results suggest that in addition to neuronal cells, various other rodent adult tissues and embryos possess a common signaling mechanism which is involved in an indirect regulation of ERK activity by VRK3-mediated VHR activity. (C) 2007 Elsevier B.V. All rights reserved.

In the eukaryotic circadian clock machinery, negative feedback repression of CLOCK (CLK) and BMAL1 transcriptional activity by PERIOD (PER) and CRYPTOCHROME (CRY) underlies the basis for 24 h rhythmic gene expression. Thus, precise regulation of the time-dependent nuclear entry of circadian repressors is crucial to generating normal circadian rhythms. Here, we sought to identify novel kinase(s) that regulate nuclear entry of mammalian CRY1 (mCRY1) with an unbiased screening using red fluorescent protein (RFP)-tagged human kinome expression plasmids in mammalian cells. Transient expression of human vaccinia-related kinase 3 (hVRK3) reduced the nuclear presence of mCRY1. hVRK3 expression also induced alterations in the subcellular localization of other core clock proteins, including mCRY2, mPER2, and BMAL1. In contrast, the subcellular localization of mCLI( was not changed. Given that singly expressed mCLI( mostly resides in the cytoplasm and that nuclear localization sequence (NLS) mutation of hVRK3 attenuated the effect of hVRK3 co-expression on subcellular localization, ectopically expressed hVRK3 presumably reduces the retention of proteins in the nucleus. Finally, downregulation of hvrk3 using siRNA reduced the amplitude and lengthened the period of the cellular bioluminescence rhythm. Taken together, these data suggest that VRK3 plays a role in setting the amplitude and period length of circadian rhythms in mammalian cells.(C) 2017 Elsevier Inc. All rights reserved.