Identification and characterization of the STIM (stromal interaction molecule) gene family: coding for a novel class of transmembrane proteins

2001 ◽  
Vol 357 (3) ◽  
pp. 673-685 ◽  
Author(s):  
Richard T. WILLIAMS ◽  
Shehnaaz S. M. MANJI ◽  
Nigel J. PARKER ◽  
Manuela S. HANCOCK ◽  
Leonie van STEKELENBURG ◽  
...  
Keyword(s):  
Single Gene ◽  
Coiled Coil ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Protein Sequencing ◽  
Ancestral Gene ◽  
Cdna Sequences ◽  
Extracellular Region ◽  
Chromosome 11P15.5

STIM1 (where STIM is stromal interaction molecule) is a candidate tumour suppressor gene that maps to human chromosome 11p15.5, a region implicated in a variety of cancers, particularly embryonal rhabdomyosarcoma. STIM1 codes for a transmembrane phosphoprotein whose structure is unrelated to that of any other known proteins. The precise pathway by which STIM1 regulates cell growth is not known. In the present study we screened gene databases for STIM1-related sequences, and have identified and characterized cDNA sequences representing a single gene in humans and other vertebrates, which we have called STIM2. We identified a single STIM homologue in Drosophila melanogaster (D-Stim) and Caenorhabditis elegans, but no homologues in yeast. STIM1, STIM2 and D-Stim have a conserved genomic organization, indicating that the vertebrate family of two STIM genes most probably arose from a single ancestral gene. The three STIM proteins each contain a single SAM (sterile α-motif) domain and an unpaired EF hand within the highly conserved extracellular region, and have coiled-coil domains that are conserved in structure and position within the cytoplasmic region. However, the STIM proteins diverge significantly within the C-terminal half of the cytoplasmic domain. Differential levels of phosphorylation appear to account for two molecular mass isoforms (105 and 115kDa) of STIM2. We demonstrate by mutation analysis and protein sequencing that human STIM2 initiates translation exclusively from a non-AUG start site in vivo. STIM2 is expressed ubiquitously in cell lines, and co-precipitates with STIM1 from cell lysates. This association into oligomers in vivo indicates a possible functional interaction between STIM1 and STIM2. The structural similarities between STIM1, STIM2 and D-STIM suggest conserved biological functions.

scholarly journals Sentinel interaction mapping – a generic approach for the functional analysis of human disease gene variants using yeast

2020 ◽  
Vol 13 (7) ◽  
pp. dmm044560
Author(s):  
Barry P. Young ◽  
Kathryn L. Post ◽  
Jesse T. Chao ◽  
Fabian Meili ◽  
Kurt Haas ◽  
...  
Keyword(s):  
Genetic Variants ◽  
Human Disease ◽  
Disease Gene ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Disease Genes ◽  
Gene Variants ◽  
Human Disease Gene ◽  
Simple Growth

ABSTRACTAdvances in sequencing technology have led to an explosion in the number of known genetic variants of human genes. A major challenge is to now determine which of these variants contribute to diseases as a result of their effect on gene function. Here, we describe a generic approach using the yeast Saccharomyces cerevisiae to quickly develop gene-specific in vivo assays that can be used to quantify the level of function of a genetic variant. Using synthetic dosage lethality screening, ‘sentinel’ yeast strains are identified that are sensitive to overexpression of a human disease gene. Variants of the gene can then be functionalized in a high-throughput fashion through simple growth assays using solid or liquid media. Sentinel interaction mapping (SIM) has the potential to create functional assays for the large majority of human disease genes that do not have a yeast orthologue. Using the tumour suppressor gene PTEN as an example, we show that SIM assays can provide a fast and economical means to screen a large number of genetic variants.

scholarly journals Sentinel Interaction Mapping (SIM) – A generic approach for the functional analysis of human disease gene variants using yeast

2020 ◽  
Author(s):  
Barry P Young ◽  
Kathryn L Post ◽  
Jesse T Chao ◽  
Fabian Meili ◽  
Kurt Haas ◽  
...  
Keyword(s):  
Genetic Variants ◽  
Human Disease ◽  
Disease Gene ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Disease Genes ◽  
Gene Variants ◽  
Human Disease Gene ◽  
Simple Growth

AbstractAdvances in sequencing technology have led to an explosion in the number of known genetic variants of human genes. A major challenge is to now determine which of these variants contribute to diseases as a result of their effect on gene function. Here we describe a generic approach using the yeast Saccharomyces cerevisiae to quickly develop gene-specific in vivo assays that can be used to quantify the level of function of a genetic variant. Using Synthetic Dosage Lethality screening, “sentinel” yeast strains are identified that are sensitive to overexpression of a human disease gene. Variants of the gene can then be functionalized in high-throughput fashion through simple growth assays using either solid or liquid media. Sentinel Interaction Mapping (SIM) has the potential to create functional assays for the large majority of human disease genes that do not have a yeast orthologue. Using the tumour suppressor gene PTEN as an example, we show that SIM assays can provide a fast and economical means to screen a large number of genetic variants.

CLL Tumours with a Deletion of 11q Form Two Functional Subgroups Based on the Mutation Status of the Remaining ATM Allele.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 710-710
Author(s):  
Belinda Austen ◽  
Maria Podinovskaia ◽  
Claire Almond ◽  
Graham Fews ◽  
Anne Gardiner ◽  
...  
Keyword(s):  
Dna Damage ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Fish Analysis ◽  
Wild Type ◽  
Atm Gene ◽  
Atm Protein ◽  
11Q Deletion

Abstract Deletions in chromosome 11q are an established prognostic marker in CLL. One copy of the ATM gene is deleted in these tumours, as detected by FISH analysis. However, it remains unclear whether the ATM gene is the main tumour suppressor gene that is accounting for the poor outcome in tumours with 11q deletions. We have recently reported that patients whose tumours have mutations in the ATM gene have an impaired overall and treatment free survival. In our large cohort of 155 patients, tumours with an ATM mutation only partly correlated with tumours with an 11q deletion. We have therefore investigated the relationship between 11q deletions and mutations in the ATM gene. Using the highly sensitive DHPLC method, we have screened the 60 ATM coding exons for mutations in a cohort of 46 tumours, all with a deletion of chromosome 11q. We have found ATM mutations in 19 tumours, indicating a prevalence of 41%. The ATM protein is vital in the cell’s response to DNA damage including that induced by chemotherapy. ATM acts upstream from p53 and defects in ATM function, like p53, lead to impaired DNA damage induced apoptosis. Furthermore, we have previously shown that loss of ATM function is associated with both in vitro and in vivo chemo-resistance. Therefore, we next assessed whether the status of the remaining ATM allele in the 11q deleted tumours affected the response to DNA damage. Firstly we induced DNA damage with irradiation and measured both the phosphorylation of ATM protein targets and the induction of p53 dependent transcription responses in representative samples. We found that 11q deleted tumours with a remaining wild type ATM allele had responses that were similar to those seen in tumours with two wild type ATM alleles. In contrast, 11q deleted tumours with a mutation in the remaining ATM allele had defective DNA damage induced responses. We then analysed the effects of in vitro treatment with Fludarabine. First we demonstrated that fludarabine induces ATM dependent phophosphorylation responses in CLL tumours. Then we analysed its effect in the two 11q deleted CLL subgroups. We showed defective phosphorylation responses to fludarabine in the tumours with a mutation in the second ATM allele, but normal responses in those with a second wild type ATM allele. In summary, we have shown that approximately 40% of CLL tumours with an 11q deletion have a mutation in their remaining ATM allele. Furthermore, we have demonstrated that the 11q deleted tumours appear to form two functional subgroups based on the presence of a mutation in the remaining ATM allele. In contrast to the subgroup with a wild type ATM allele, CLL tumours with a mutant ATM allele have defective in vitro responses to DNA damage with both irradiation and fludarabine. We expect that the functional differences between the two 11q deleted subsets will translate into differences in clinical outcome.

scholarly journals Low GAS5 expression may predict poor survival and cisplatin resistance in cervical cancer

2020 ◽  
Vol 11 (7) ◽  
Author(s):  
Xingyu Fang ◽  
Guanglei Zhong ◽  
Yuhan Wang ◽  
Zhongqiu Lin ◽  
Rongchun Lin ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Animal Studies ◽  
Cisplatin Resistance ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Luciferase Reporter ◽  
Poor Overall Survival ◽  
Cisplatin Sensitivity

Abstract Cisplatin resistance is a major challenge in cervical cancer (CC) chemotherapy. Growth arrest‐specific 5 (GAS5) has been reported to be a tumour suppressor gene in CC. However, the mechanism of GAS5 in chemoresistance remains undetermined. Our research evaluated GAS5 expression in normal and CC tissues by qPCR and in situ hybridization (ISH). Statistical analysis was conducted to analyse the association of GAS5 expression with survival. Biochemical methods were used to screen upstream and downstream regulators of GAS5. Then, interactions were confirmed by ChIP, RNA pull-down, RNA immunoprecipitation (RIP), dual-luciferase reporter and real-time PCR assays. The cisplatin sensitivity of GAS5-overexpressing CC cells was demonstrated in vitro and in vivo. The results showed that low GAS5 expression was correlated with poor overall survival. Mechanistically, GAS5 was transcriptionally modulated by P-STAT3 and served as a competing endogenous RNA (ceRNA) of miR-21 to indirectly affect cisplatin sensitivity through PDCD4 regulation in CC cells. Animal studies confirmed that GAS5 enhanced cisplatin sensitivity and promoted PDCD4 expression in vivo. GAS5 was regulated by P-STAT3 and affected the sensitivity of CC to cisplatin-based chemotherapy through the miR-21/PDCD4 axis. This result may provide new insight into cisplatin-based therapy.

Functional analysis of the tumour suppressor gene PTEN in murine B cells and keratinocytes

2004 ◽  
Vol 32 (2) ◽  
pp. 362-365 ◽  
Author(s):  
A. Suzuki ◽  
T. Sasaki ◽  
T.W. Mak ◽  
T. Nakano
Keyword(s):  
B Cells ◽  
Cytidine Deaminase ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Mutant Mice ◽  
Specific Gene ◽  
Marginal Zone B Cells ◽  
Specific Mutation ◽  
Phosphoinositide 3 Kinase

To investigate the roles of the PTEN (phosphatase and tensin homologue deleted from chromosome 10)/PI3K (phosphoinositide 3-kinase) signalling pathway in vivo, we have generated a series of mutant mice with null or tissue-specific gene-targeted deletions of Pten. Here we present our investigations of Pten function in B cells and keratinocytes in mice. Mice with a B cell-specific mutation of Pten showed increased serum autoantibodies and elevated numbers of B1a cells. Among conventional B (B2) cells in mutant spleens, numbers of marginal zone B cells were significantly increased, while those of follicular B cells were reciprocally decreased. Immunoglobulin class switch recombination was defective and associated with impaired induction of activation-induced cytidine deaminase. Mice with a keratinocyte-specific mutation of Pten exhibited epidermal hyperplasia, hyperkeratosis and accelerated skin morphogenesis. Within 3 weeks of birth, 90% of these animals died of malnutrition, possibly caused by hyperkeratosis of the oesophageal epithelia. Surviving mutant mice developed spontaneous skin tumours within 8.5 months of birth, and chemical treatment accelerated the onset of tumours. Our data show that PTEN is an important regulator in B cells and keratinocytes.

scholarly journals Epigenetic silencing of the XAF1 gene is mediated by the loss of CTCF binding

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Georgina Victoria-Acosta ◽  
Karla Vazquez-Santillan ◽  
Luis Jimenez-Hernandez ◽  
Laura Muñoz-Galindo ◽  
Vilma Maldonado ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Cell Cycle Progression ◽  
Molecular Mechanisms ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Epigenetic Silencing ◽  
Ctcf Binding ◽  
Open Chromatin ◽  
Cpg Dinucleotide

Abstract XAF1 is a tumour suppressor gene that compromises cell viability by modulating different cellular events such as mitosis, cell cycle progression and apoptosis. In cancer, the XAF1 gene is commonly silenced by CpG-dinucleotide hypermethylation of its promoter. DNA demethylating agents induce transcriptional reactivation of XAF1, sensitizing cancer cells to therapy. The molecular mechanisms that mediate promoter CpG methylation have not been previously studied. Here, we demonstrate that CTCF interacts with the XAF1 promoter in vivo in a methylation-sensitive manner. By transgene assays, we demonstrate that CTCF mediates the open-chromatin configuration of the XAF1 promoter, inhibiting both CpG-dinucleotide methylation and repressive histone posttranslational modifications. In addition, the absence of CTCF in the XAF1 promoter inhibits transcriptional activation induced by well-known apoptosis activators. We report for the first time that epigenetic silencing of the XAF1 gene is a consequence of the loss of CTCF binding.

A mouse homologue of the Drosophila tumour-suppressor gene l(2)gl controlled by Hox-C8 in vivo

Nature ◽  
1993 ◽  
Vol 365 (6441) ◽  
pp. 69-72 ◽  
Author(s):  
Daihachiro Tomotsune ◽  
Hiroki Shoji ◽  
Yoshio Wakamatsu ◽  
Hisato Kondoh ◽  
Naoki Takahashi
Keyword(s):  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Tumour Suppressor ◽  
Mouse Homologue

scholarly journals Identification of a carbonic anhydrase-like domain in the extracellular region of RPTP gamma defines a new subfamily of receptor tyrosine phosphatases.

1993 ◽  
Vol 13 (3) ◽  
pp. 1497-1506 ◽  
Author(s):  
G Barnea ◽  
O Silvennoinen ◽  
B Shaanan ◽  
A M Honegger ◽  
P D Canoll ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Transmembrane Domain ◽  
Tyrosine Phosphatase ◽  
Extracellular Domain ◽  
Suppressor Gene ◽  
Tyrosine Phosphatases ◽  
Candidate Tumor Suppressor Gene ◽  
Extracellular Region ◽  
Receptor Tyrosine Phosphatases

The tyrosine phosphatase RPTP gamma is a candidate tumor suppressor gene since it is located on human chromosome 3p14.2-p21 in a region frequently deleted in certain types of renal and lung carcinomas. In order to evaluate its oncogenic potential and to explore its normal in vivo functions, we have isolated cDNAs and deduced the complete sequences of both human and murine RPTP gamma. The murine RPTP gamma gene has been localized to chromosome 14 to a region syntenic to the location of the human gene. Northern (RNA) blot analysis reveals the presence of two major transcripts of 5.5 and 8.5 kb in a variety of murine tissues. In situ hybridization analysis reveals that RPTP gamma mRNA is expressed in specific regions of the brain and that the localization of RPTP gamma changes during brain development. RPTP gamma is composed of a putative extracellular domain, a single transmembrane domain, and a cytoplasmic portion with two tandem catalytic tyrosine phosphatase domains. The extracellular domain contains a stretch of 266 amino acids with striking homology to the zinc-containing enzyme carbonic anhydrase (CAH), indicating that RPTP gamma and RPTP beta (HPTP zeta) represent a subfamily of receptor tyrosine phosphatases. We have constructed a model for the CAH-like domain of RPTP gamma based upon the crystal structure of CAH. It appears that 11 of the 19 residues that form the active site of CAH are conserved in RPTP gamma. Yet only one of the three His residues that ligate the zinc atom and are required for catalytic activity is conserved. On the basis of this model we propose that the CAH-like domain of RPTP gamma may have a function other than catalysis of hydration of metabolic CO2.

scholarly journals Glucocorticoid receptors in lung cancer: new perspectives

2016 ◽  
Vol 229 (1) ◽  
pp. R17-R28 ◽  
Author(s):  
Kerryn M Taylor ◽  
David W Ray ◽  
Paula Sommer
Keyword(s):  
Lung Cancer ◽  
Lung Diseases ◽  
Glucocorticoid Receptors ◽  
Ex Vivo ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Lung Cancers ◽  
Synthetic Glucocorticoids

Proper expression of the glucocorticoid receptor (GR) plays an essential role in the development of the lung. GR expression and signalling in the lung is manipulated by administration of synthetic glucocorticoids (Gcs) for the treatment of neonatal, childhood and adult lung diseases. In lung cancers, Gcs are also commonly used as co-treatment during chemotherapy. This review summarises the effect of Gc monotherapy and co-therapy on lung cancers in vitro, in mouse models of lung cancer, in xenograft, ex vivo and in vivo. The disparity between the effects of pre-clinical and in vivo Gc therapy is commented on in light of the recent discovery of GR as a novel tumour suppressor gene.

844 Blocking expression of the tumour suppressor gene, p63 inhibits in-vivo carcinogenesis in prostate cancer

2014 ◽  
Vol 13 (1) ◽  
pp. e844
Author(s):  
R. Beekharry ◽  
W. Bansema ◽  
P. Berry ◽  
H. Walker ◽  
A. Nicholl ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Suppressor Gene ◽  
Tumour Suppressor Gene ◽  
Tumour Suppressor
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