Two short sequences have positive effects on the human p27 gene transcription

Ito, Emi; Iwahashi, Yuki; Yanagisawa, Yuka; Suzuki, Yutaka; Sugano, Sumio; Yuasa, Yasuhito; Maruyama, Kazuo

doi:10.1016/s0378-1119(99)00022-0

Cited by 19 publications

(22 citation statements)

References 26 publications

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“…We cloned the minimal regulatory region of the CDKN1B gene that has been shown to be able to efficiently drive transcription of a downstream reporter gene (29,30,31). DNA fragments from the 5 0 region of the CDKN1B gene between nucleotides K575 and K1 were generated through PCR using genomic DNA of a donor sample with forward 5 0 -GGTACC-GGTACCCCACCTTAAGGCCGCGCTCG-3 0 and reverse 5 0 -AAGCTTA AGCTTCTTTCTCCCGGGTCTGCACG-3 0 primers containing KpnI and HindIII sites respectively.…”

Section: Constructsmentioning

confidence: 99%

“…Transcription from CDKN1B is initiated predominantly from a single site which is conserved in the human and mouse genes (29,30,31). Initiation at this site produces a 5 0 -UTR of 472 nucleotides in the human CDKN1B mRNA and 502 nucleotides in the mouse Cdkn1B mRNA.…”

Section: Functional Effects Of Thementioning

confidence: 99%

“…Initiation at this site produces a 5 0 -UTR of 472 nucleotides in the human CDKN1B mRNA and 502 nucleotides in the mouse Cdkn1B mRNA. In addition, several minor transcription start sites were identified for both the mouse and human genes (29,30,31). In addition to the capability to drive transcription of a downstream reporter gene, the 5 0 -UTR region of human CDKN1B contains a U-rich element that is involved in regulating CDKN1B mRNA stability and/or translation efficacy (33).…”

Section: Functional Effects Of Thementioning

confidence: 99%

“…In fact, several mRNA-binding proteins were found to interact with the CDKN1B 5 0 -UTR and to regulate its expression (34). For this reason, to investigate whether the deleted 5 0 -UTR sequence detected in the patient ID7 affected the expression of CDKN1B mRNA and/or protein, we decided to clone the minimal regulatory region of the CDKN1B gene that has been shown to efficiently drive transcription of a downstream reporter gene (30,31,35,36) and measure the effects on transcription exerted by the mutant allele. For this purpose, the 575 bp sequence in the CDKN1B 5 0 -UTR was PCR amplified from wild-type or mutant genomic DNAs and inserted upstream of the firefly luciferase reporter gene in the promoterless pGL3 vector, generating wild-type (5 0 -UTR-WT) and deleted (5 0 -UTR-MUT) constructs (Fig.…”

Section: Functional Effects Of Thementioning

confidence: 99%

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Functional characterization of a rare germline mutation in the gene encoding the cyclin-dependent kinase inhibitor p27Kip1 (CDKN1B) in a Spanish patient with multiple endocrine neoplasia-like phenotype

Malanga¹,

Gisi²,

Riccardi³

et al. 2012

European Journal of Endocrinology

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Objective: The aim of this study was to investigate the presence of germline mutations in the CDKN1B gene that encodes the cyclin-dependent kinase (Cdk) inhibitor p27 in multiple endocrine neoplasia 1 (MEN1)-like Spanish index patients. The CDKN1B gene has recently been identified as a tumor susceptibility gene for MEN4, with six germline mutations reported so far in patients with a MEN-like phenotype but negative for MEN1 mutations. Design and methods: Fifteen Spanish index cases with MEN-like symptoms were screened for mutations in the CDKN1B gene and the mutant variant was studied functionally by transcription/translation assays in vitro and in transiently transfected HeLa cells. Results: We report the identification of a heterozygous GAGA deletion in the 5 0 -UTR of CDKN1B, NM_004064.3:c.-32_-29del, in a patient affected by gastric carcinoid tumor and hyperparathyroidism. This deletion falls inside the region that is responsible for CDKN1B transcription and is predicted to destroy a secondary stem and loop structure that includes the GAGAGA element responsible for ribosome recruitment. Accordingly, in vitro studies of coupled transcription/translation assays and transient transfection in HeLa cells showed that the GAGA deletion in the CDKN1B 5 0 -UTR significantly impairs the transcription of downstream reporter luciferase (of w40-60%) and, possibly, the translation of the corresponding mRNA. This mutation was associated with a significant reduction in the amount of CDKN1B mRNA in peripheral blood leukocytes from the patient, as demonstrated by quantitative real-time PCR. Conclusions: Our results confirm that germline CDKN1B mutations may predispose to a human MEN4 condition and add novel evidence that alteration in the transcription/translation rate of CDKN1B mRNA might be the mechanism implicated in tumor susceptibility.

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Section: Constructsmentioning

confidence: 99%

Section: Functional Effects Of Thementioning

confidence: 99%

Section: Functional Effects Of Thementioning

confidence: 99%

Section: Functional Effects Of Thementioning

confidence: 99%

See 2 more Smart Citations

Functional characterization of a rare germline mutation in the gene encoding the cyclin-dependent kinase inhibitor p27Kip1 (CDKN1B) in a Spanish patient with multiple endocrine neoplasia-like phenotype

Malanga¹,

Gisi²,

Riccardi³

et al. 2012

European Journal of Endocrinology

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“…pHsMCM5-Luc(-1384), pHsCdc6-Luc(-570), p27 Kip1 reporters pGL3-844-S, pGL3-635-S and pGL3-499-S, expression vectors for E2F1 through E2F3, and b-galactosidase expression vector pCMV-b-gal have been described (Ohtani et al, 1998(Ohtani et al, , 1999Ito et al, 1999). pGL3-814-S and pGL3-788-S were generated by removing NheI-Bpu10I and NheI-ApaI region from pGL3-844-S, respectively.…”

Section: Plasmids and Reporter Assaymentioning

confidence: 99%

Identification of novel E2F1 target genes regulated in cell cycle-dependent and independent manners

et al. 2005

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The transcription factor E2F mediates cell cycle-dependent expression of genes important for cell proliferation in response to growth stimulation. To further understand the role of E2F, we utilized a sensitive subtraction method to explore new E2F1 targets, which are expressed at low levels and might have been unrecognized in previous studies. We identified 33 new E2F1-inducible genes, including checkpoint genes Claspin and Rad51ap1, and four genes with unknown function required for cell cycle progression. Moreover, we found three groups of E2F1-inducible genes that were not induced by growth stimulation. At least, two groups of genes were directly induced by E2F1, indicating that E2F1 can regulate expression of genes not induced during the cell cycle. One included Neogenin, WASF1 and SGEF genes, which may have a role in differentiation or development. The other was the cyclin-dependent kinase inhibitor p27 Kip1 , which was involved in suppression of inappropriate cell cycle progression induced by deregulated E2F. E2F1-responsive regions of these genes were located more upstream than those of typical E2F targets and did not have typical E2F sites. These results indicate that there are groups of E2F1 targets, which are regulated in a distinct manner from that of typical E2F targets.

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IGF-II Enhances Trichostatin A-Induced TGFβ1 and p21Waf1,Cip1,Sdi1 Expression in Hep3B Cells

Gray

Yakovleva

Hartmann

et al. 1999

Experimental Cell Research

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Two short sequences have positive effects on the human p27 gene transcription

Cited by 19 publications

References 26 publications

Functional characterization of a rare germline mutation in the gene encoding the cyclin-dependent kinase inhibitor p27Kip1 (CDKN1B) in a Spanish patient with multiple endocrine neoplasia-like phenotype

Functional characterization of a rare germline mutation in the gene encoding the cyclin-dependent kinase inhibitor p27Kip1 (CDKN1B) in a Spanish patient with multiple endocrine neoplasia-like phenotype

Identification of novel E2F1 target genes regulated in cell cycle-dependent and independent manners

IGF-II Enhances Trichostatin A-Induced TGFβ1 and p21Waf1,Cip1,Sdi1 Expression in Hep3B Cells

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