Molecular Characterization of Human Zyxin

Macalma, Teresita; Otte, Jürgen; Hensler, Mary E.; Bockholt, Susanne M.; Louis, Heather A.; Kalff-Suske, Martha; Grzeschik, K.-H.; Ahe, Dietmar von der; Beckerle, Mary C.

doi:10.1074/jbc.271.49.31470

Cited by 109 publications

(118 citation statements)

References 54 publications

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“…Several other proteins can interact with HIPK1 through its p53-binding domain (Y.L. and M.D., unpublished data), including Hic5 (27), UBC9 (28), and zyxin (29). Interactions with Hic5 and zyxin are particularly interesting because these proteins are recruited into focal adhesions and provide a link with the extracellular matrix (focal adhesion complex; refs.…”

Section: Discussionmentioning

confidence: 99%

Characterization of cells and gene-targeted mice deficient for the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1)

Kondo

Debbas

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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The tumor suppressor p53 is regulated in part by binding to cellular proteins. We used p53 as bait in the yeast two-hybrid system and isolated homeodomain-interacting protein kinase 1 (HIPK1) as a p53-binding protein. Deletion analysis showed that amino acids 100 -370 of p53 and amino acids 885-1093 of HIPK1 were sufficient for HIPK1-p53 interaction. HIPK1 was capable of autophosphorylation and specific serine phosphorylation of p53. The HIPK1 gene was highly expressed in human breast cancer cell lines and oncogenically transformed mouse embryonic fibroblasts. HIPK1 was localized to human chromosome band 1p13, a site frequently altered in cancers. Gene-targeted HIPK1؊͞؊ mice were grossly normal but oncogenically transformed HIPK1 ؊͞؊ mouse embryonic fibroblasts exhibited reduced transcription of Mdm2 and were more susceptible than transformed HIPK1؉͞؉ cells to apoptosis induced by DNA damage. Carcinogen-treated HIPK1 ؊͞؊ mice developed fewer and smaller skin tumors than HIPK1؉͞؉ mice. HIPK1 may thus play a role in tumorigenesis, perhaps by means of the regulation of p53 and͞or Mdm2. T he most frequently mutated gene associated with human cancers is p53 (1, 2). In response to cellular stresses such as DNA damage and oncogene expression, p53 promotes cell-cycle arrest or apoptosis (3, 4). p53 stabilization and activity are regulated by conformational changes (3) and posttranslational modification by phosphorylation (5). However, the kinase(s) relevant for phosphorylation of p53 in vivo remain obscure. p53 activity is also regulated by interaction with cellular molecules such as the oncogenic protein Mdm2 (2). Overexpression of Mdm-2 in certain tumors results in the inactivation and degradation of p53 (6-10). We set out to identify proteins that interact with p53 and to examine the expression of these molecules in transformed cells. We isolated the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1), previously identified as a homeodomain-interacting protein (11), and analyzed p53-HIPK1 interaction by various means. The physiological functions of HIPK1 were characterized by using gene-targeted HIPK1-deficient mice. Our in vivo and in vitro characterizations of HIPK1 suggest that HIPK1 is a modulator of p53 activity that can promote oncogenesis. MethodsYeast Two-Hybrid Screening. A human p53 cDNA (amino acids 71-393; ref. 12) was subcloned in-frame into pGBT9 (GAL4 DNA-binding domain vector) and used to screen cDNA libraries constructed by using pGAD424 (GAL4-activation-domain plasmid; ref. 13) in yeast Y2H strain SFY526. The libraries were derived from rat embryo fibroblasts transformed with E1A ϩ Ras ϩ mutant p53 R273H, and a breast cancer cell line. ␤-Galactosidase activities were measured by using the GalactoLight reporter assay kit (Tropix, Bedford, MA). The cDNA plasmids from ␤-galactosidase-positive yeast were rescued, retested in yeast, and sequenced.Interaction of HIPK1 with p53 in 293 Cells. The SalI͞NotI HIPK1 fragment corresponding to amino acids 678-1149 was ligated in-frame with either ...

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Section: Discussionmentioning

confidence: 99%

Characterization of cells and gene-targeted mice deficient for the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1)

Kondo

Debbas

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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“…Encodes a LIM domain protein localized at focal contacts in adherent erythroleukemia cells (Macalma, 1996).…”

Section: All > Aml Hg161macmarcksmentioning

confidence: 99%

Untitled

Guyon¹,

Weston²,

Barnhill³

et al. 2002

Machine Learning

7,759

1,340

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Abstract. DNA micro-arrays now permit scientists to screen thousands of genes simultaneously and determine whether those genes are active, hyperactive or silent in normal or cancerous tissue. Because these new micro-array devices generate bewildering amounts of raw data, new analytical methods must be developed to sort out whether cancer tissues have distinctive signatures of gene expression over normal tissues or other types of cancer tissues.In this paper, we address the problem of selection of a small subset of genes from broad patterns of gene expression data, recorded on DNA micro-arrays. Using available training examples from cancer and normal patients, we build a classifier suitable for genetic diagnosis, as well as drug discovery. Previous attempts to address this problem select genes with correlation techniques. We propose a new method of gene selection utilizing Support Vector Machine methods based on Recursive Feature Elimination (RFE). We demonstrate experimentally that the genes selected by our techniques yield better classification performance and are biologically relevant to cancer.In contrast with the baseline method, our method eliminates gene redundancy automatically and yields better and more compact gene subsets. In patients with leukemia our method discovered 2 genes that yield zero leaveone-out error, while 64 genes are necessary for the baseline method to get the best result (one leave-one-out error). In the colon cancer database, using only 4 genes our method is 98% accurate, while the baseline method is only 86% accurate.

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“…Group 2 proteins include cystein-rich intestinal protein (CRIP; accession number, M33146) (Liebhaber et al 1990) and cystein-rich protein (CRP; accession number, M76375) (Wang et al 1992), which contain a short additional conserved motif after the LIM domain. Group 3 proteins include Ril (accession number, X76454) (Kiess et al 1995), paxillin (accession number, U14588) (Salgia et al 1995), zyxin (accession number, X94991) (Macalma et al 1996), Enigma (accession number, L35240) (Wu and Gill 1994), and ENH (accession number, U48247) (Kuroda et al 1996). Although this group is heterogeneous, the LIM domains in this group are more closely related to each other than to those in groups 1 and 2.…”

Section: Introductionmentioning

confidence: 99%

Isolation, tissue expression, and chromosomal assignment of a human LIM protein gene, showing homology to rat Enigma homologue (ENH)

Ueki

Seki²,

Yano

et al. 1999

J Hum Genet

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Rat ENH (Enigma homolog) is a LIM domain protein that associates with protein kinase C in an isoformspecific manner. We have identified a human cDNA which shares a significant sequence homology with rat ENH. The isolated cDNA clone, designated human ENH (hENH), was 3287 bp in length and encoded a predicted protein of 596 amino acids which had 88% overall identity to rat ENH protein. Northern blot analysis revealed that 1.9 kb of the hENH messenger RNA was predominantly expressed in heart and skeletal muscle, while 5.6 kb of the hENH messenger RNA was ubiquitously expressed in various human tissues. The chromosomal location of the gene was determined on chromosome 4q22 region, between markers WI-2900 and WI-3273, by polymerase chain reaction (PCR)-based analyses using both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel.

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Molecular Characterization of Human Zyxin

Cited by 109 publications

References 54 publications

Characterization of cells and gene-targeted mice deficient for the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1)

Characterization of cells and gene-targeted mice deficient for the p53-binding kinase homeodomain-interacting protein kinase 1 (HIPK1)

Untitled

Isolation, tissue expression, and chromosomal assignment of a human LIM protein gene, showing homology to rat Enigma homologue (ENH)

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