Functional characterization of human PFTK1 as a cyclin-dependent kinase

Shu, Fang; Lv, Shun; Yan, Qi; Ma, Xiaoliang; Wang, Xin; Peng, Xiaozhong; Luo, Ying; Xu, Bing-e; Sun, Xiao‐Feng; Wu, Jun

doi:10.1073/pnas.0703327104

Cited by 58 publications

(50 citation statements)

References 31 publications

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“…Pftk1 (Pftaire1) is a cdc2-related protein kinase involved in cell cycle progression, proliferation, migration and motility Shu et al, 2007a). The fact that higher amounts of myocilin lead to changes in gene expression in the retina suggests that myocilin might also have a direct or indirect function for retinal cells in the normal eye.…”

Section: Discussionmentioning

confidence: 99%

Elevated amounts of myocilin in the aqueous humor of transgenic mice cause significant changes in ocular gene expression

Paper

Kroeber

Heersink

et al. 2008

Experimental Eye Research

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Myocilin is a 55-57kDa secreted glycoprotein and member of the olfactomedin family, which is mutated in some forms of primary open-angle glaucoma. To assess the effects of elevated amounts of myocilin on aqueous humor outflow dynamics in an in vivo system, transgenic betaB1-crystallin-MYOC mice have been developed that strongly overexpress myocilin in their eyes. The transgenic overexpression of myocilin results in an almost five-fold increase of secreted normal myocilin in the aqueous humor of betaB1-crystallin-MYOC mice. In the present study, we wanted to use betaB1-crystallin-MYOC as a tool to identify the response of ocular tissues to the presence of higher than normal amounts of myocilin, and to identify changes in gene expression that could help to shed light on the functional in vivo properties of myocilin. RNA was isolated from ocular tissues of betaB1-crystallin-MYOC mice and wild-type littermates. Changes in gene expression were determined by hybridization of gene microarrays and confirmed by real time RT-PCR and Western blotting. The expression of genes that had been found to be differentially regulated in betaB1-crystallin-MYOC mice was further analyzed in cultured human trabecular meshwork (HTM) cells treated with recombinant myocilin. Although betaB1-crystallin-MYOC mice do not have an obvious phenotype, a statistically significant up- and downregulation of several distinct genes was found when compared to gene expression in wild-type littermates. Among the genes that were found to be differentially regulated were Wasl, Ceacam1, and Spon2, which are involved in cell adhesion and cell-matrix interactions. Differences in expression were also found for Six1 which encodes for a transcription factor, and for Pftk1 whose gene product is a cdc2-related protein kinase. The expression of these genes was also found to be regulated in vitro in HTM cells treated with recombinant myocilin. Substantially higher amounts in ocular tissues of betaB1-crystallin-MYOC mice were found for connexin 46 and alphaB-crystallin. In addition, several genes that encode for olfactomedin proteins showed distinct changes in expression. Olfml3 was significantly downregulated, while Lphn1, Lphn2, and Lphn3 were significantly upregulated. Our findings support a role for myocilin in modulating cellular adhesion, and suggest functional processes that involve other proteins of the olfactomedin family.

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Section: Discussionmentioning

confidence: 99%

Elevated amounts of myocilin in the aqueous humor of transgenic mice cause significant changes in ocular gene expression

Paper

Kroeber

Heersink

et al. 2008

Experimental Eye Research

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“…A potential role for Pftk1 in cell cycle progression and cell proliferation has been suggested. 32 Pftk1 is also expressed in postmitotic cells 33 and thus may have functions beyond the cell cycle. Pftk1 mRNA expression does not follow the same kinetics as CD11c and DC-STAMP in DCs ( Figure 7B) as it is also present in bone marrow cells (as suggested by public microarray data; GEO profiles, National Center for Biotechnology Information).…”

Section: Discussionmentioning

confidence: 99%

Comparative promoter analysis in vivo: identification of a dendritic cell-specific promoter module

Edelmann

Nelson

Brocker

2011

Blood

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Dendritic cells (DCs) are important immune cells. This study focused on transcriptional networks active in murine DCs, but DCs are difficult to study using conventional molecular techniques. Therefore, comparative promoter analysis was used to identify evolutionarily conserved features between the murine CD11c and DC-STAMP promoters. A promoter framework consisting of 4 transcription factor binding sites was identified that included signal transducer and activator of transcription, homeodomain transcription factors, and 2 members of the Brn POU domain factors family. This promoter module was functionally verified by in vivo promoter analysis and site-directed mutagenesis. Hematopoietic stem cells were engineered by lentiviral vectors and expression of green fluorescent protein reporter was monitored in primary hematopoietic cell types that develop without further manipulation in irradiated recipient mice. The verified promoter module was then modeled and used in a bioinformatics-based search for other potential coregulated genes in murine DCs. A promoter database search identified 2 additional genes, Ppef2 and Pftk1, which have a similar promoter organization and are preferentially expressed in murine DCs. The results define a regulatory network linked to development of murine DCs. (Blood. 2011;118(11):e40-e49) IntroductionDendritic cells (DCs) play key roles in shaping the immune response. 1,2 They are highly specialized in uptake and presentation of antigen. In their steady state, DCs migrate from peripheral tissues to the draining lymph node and present self-antigen to naive T cells, inducing tolerance. On encounter of pathogen, DCs change their phenotype (undergo maturation), which leads to a more pronounced migratory behavior and an increase in stimulatory capacity. 3 Plasticity is a pervasive feature of DC biology. DC subsets from different tissues show differential morphology, phenotypes, and functions. An additional distinction can also be made between conventional DCs (cDCs) and plasmacytoid DCs (pDCs), which are mainly involved in viral immune responses. cDCs also include a subset (CD8 ϩ in mice) specialized in antigen cross-presentation. 4 Networks of genes linked by common control mechanisms may help orchestrate homeostasis, as well as the response of DCs to external stimuli. The prediction and analysis of these networks represent a major challenge in understanding immunology. Transcriptional mechanisms have been very difficult to study in DCs because of their propensity to quickly change phenotypes in response to physical manipulation and their limited numbers. Gene expression is controlled by a series of processes that include locus control regions, chromatin rearrangement, methylation, gene enhancers, etc. These regulatory events ultimately end at the promoter level to effect gene transcription. Promoters integrate this information to achieve the tissue and signal specific control of gene expression required for complex biologic processes, such as DC biology and function.Within promoters, transcri...

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“…To analyze protein interactions in vitro, approximately 1 × 10 7 HEK293 cells (Cell Bank, Chinese Academy of Sciences) were harvested ~24-36 h after transfection and lysed in 1 ml of RIPA buffer (50 mM Tris-Cl (pH 7.5), 150 mM NaCl, 10% glycerol (w/v), 0.5% sodium deoxycholate, 0.1% SDS, 1% NP-40, 1 mM EDTA and protease inhibitors). The resulting lysates were subjected to immunoprecipitation with antibodies directed against the epitope tag as described previously 45 Yeast two-hybrid assays. Ret intracellular domain (ICD)-interacting proteins were identified using yeast two-hybrid screening, as described previously 45 .…”

Section: Competing Financial Interestsmentioning

confidence: 99%

“…The resulting lysates were subjected to immunoprecipitation with antibodies directed against the epitope tag as described previously 45 Yeast two-hybrid assays. Ret intracellular domain (ICD)-interacting proteins were identified using yeast two-hybrid screening, as described previously 45 . The Ret ICD and Vav2 cDNAs were subcloned into the yeast expression vectors pGBT7 and pACT2, respectively (Clontech).…”

Section: Competing Financial Interestsmentioning

confidence: 99%

Identification of a Vav2-dependent mechanism for GDNF/Ret control of mesolimbic DAT trafficking

Zhu

Zhao

et al. 2015

Nat Neurosci

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Dopamine (DA) homeostasis is essential for a variety of brain activities. Dopamine transporter (DAT)-mediated DA reuptake is one of the most critical mechanisms for normal DA homeostasis. However, the molecular mechanisms underlying the regulation of DAT activity in the brain remain poorly understood. Here we show that the Rho-family guanine nucleotide exchange factor protein Vav2 is required for DAT cell surface expression and transporter activity modulated by glial cell line-derived neurotrophic factor (GDNF) and its cognate receptor Ret. Mice deficient in either Vav2 or Ret displayed elevated DAT activity, which was accompanied by an increase in intracellular DA selectively in the nucleus accumbens. Vav2(-/-) mice exposed to cocaine showed reduced DAT activity and diminished behavioral cocaine response. Our data demonstrate that Vav2 is a determinant of DAT trafficking in vivo and contributes to the maintenance of DA homeostasis in limbic DA neuron terminals.

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Functional characterization of human PFTK1 as a cyclin-dependent kinase

Cited by 58 publications

References 31 publications

Elevated amounts of myocilin in the aqueous humor of transgenic mice cause significant changes in ocular gene expression

Elevated amounts of myocilin in the aqueous humor of transgenic mice cause significant changes in ocular gene expression

Comparative promoter analysis in vivo: identification of a dendritic cell-specific promoter module

Identification of a Vav2-dependent mechanism for GDNF/Ret control of mesolimbic DAT trafficking

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