Properties of a novel gene isolated from a Hodgkin's disease cell line that is expressed early during lymphoid cell activation

We have isolated a novel 667-bp cDNA clone, designated epag, from a Hodgkin's-disease cell line-derived library that is expressed in association with T cell activation and which is not related to any known gene family. By using reverse transcription/PCR, we have demonstrated that epag mRNA is e...

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Veröffentlicht in:The Journal of immunology (1950) 1994-03, Vol.152 (5), p.2229-2240
Hauptverfasser: Bennett, JS, Tredway, TL, Dizikes, GJ, Nawrocki, JF
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container_title The Journal of immunology (1950)
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creator Bennett, JS
Tredway, TL
Dizikes, GJ
Nawrocki, JF
description We have isolated a novel 667-bp cDNA clone, designated epag, from a Hodgkin's-disease cell line-derived library that is expressed in association with T cell activation and which is not related to any known gene family. By using reverse transcription/PCR, we have demonstrated that epag mRNA is expressed as early as 1 h after stimulation of normal PBMCs with anti-CD3. The levels of mRNA peaked by 4 h, and no expression was detectable by 12 h postactivation or in resting cells incubated in culture without activation. Expression of epag was also detected in PMA- and PHA-stimulated, but not in nonstimulated Jurkat cells, and overall its expression in transformed cell lines of hemopoietic origin is highly restricted. Sequence analysis of multiple independent cDNA clones showed that epag expressed in the Hodgkin's-disease cell line L428 is identical to the gene expressed in normal activated PBMC. Epag expression was detected by reverse transcription/PCR in RNA preparations made from various normal nonlymphoid tissues. Computer analysis of the sequence identified an open reading frame encoding a putative protein of 13.2 kDa initiating at a CUG translational codon. In vitro translation and Western blot analysis with anti-peptide serum supported this analysis. We hypothesize that epag functions as an early signal that helps mediate the activation of T cells.
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By using reverse transcription/PCR, we have demonstrated that epag mRNA is expressed as early as 1 h after stimulation of normal PBMCs with anti-CD3. The levels of mRNA peaked by 4 h, and no expression was detectable by 12 h postactivation or in resting cells incubated in culture without activation. Expression of epag was also detected in PMA- and PHA-stimulated, but not in nonstimulated Jurkat cells, and overall its expression in transformed cell lines of hemopoietic origin is highly restricted. Sequence analysis of multiple independent cDNA clones showed that epag expressed in the Hodgkin's-disease cell line L428 is identical to the gene expressed in normal activated PBMC. Epag expression was detected by reverse transcription/PCR in RNA preparations made from various normal nonlymphoid tissues. Computer analysis of the sequence identified an open reading frame encoding a putative protein of 13.2 kDa initiating at a CUG translational codon. 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Psychology</subject><subject>Gene Expression</subject><subject>GENE REGULATION</subject><subject>genes</subject><subject>Genes. Genome</subject><subject>Hodgkin Disease - genetics</subject><subject>Hodgkin Disease - immunology</subject><subject>Hodgkin's disease</subject><subject>Humans</subject><subject>HYBRIDIZATION</subject><subject>LEUKOCYTES</subject><subject>Lymphocyte Activation - genetics</subject><subject>LYMPHOCYTES</subject><subject>lymphocytes T</subject><subject>man</subject><subject>MATERIALS</subject><subject>METABOLIC ACTIVATION</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>nucleotide sequence</subject><subject>Open Reading Frames</subject><subject>prediction</subject><subject>SOMATIC CELLS</subject><subject>STRUCTURAL CHEMICAL ANALYSIS 550400 -- Genetics</subject><subject>T-Lymphocytes - immunology</subject><subject>Tumor Cells, Cultured - immunology</subject><issn>0022-1767</issn><issn>1550-6606</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU-LFDEQxYMo67j6CUQIIu6px_zrdPdRFnWFBT3oOWSS6ums6c6YpHeci5_dND0u3jwV1Pu9VwUPoZeUbAUR3bs7N47zFPyW1mxbbxlj3SO0oXVNKimJfIw2hDBW0UY2T9GzlO4IIZIwcYEuWso54XKDfn-N4QAxO0g49FjjKdyDx3uYALsUvM5gcR_DWKSbYPc_3HSVsHUJdAJswHvsXWHzoHMxYPh1iJBSMYGO_oTtHN20x_40Hobg7OrQJrt7nV2YnqMnvfYJXpznJfr-8cO365vq9sunz9fvbysjpMhVLSSnnLVM2B3tGmKpIFoa2lnWl7WwHd-ZhlupW2lk3bdAKW-hkztGTC0bfoler7khZaeScRnMYMI0gcmqIaTjoi3Q2xU6xPBzhpTV6NLysJ4gzEk1UhDCO_ZfkMq2bRq-nOUraGJIKUKvDtGNOp4UJWrpUP3tUJUOVa2WDovr1Tl-3o1gHzzn0or-5qzrZLTvo56MSw-YILSrxfLl1YoNbj8cXQSVRu19CaXqeDz-c_AP_dS0Aw</recordid><startdate>19940301</startdate><enddate>19940301</enddate><creator>Bennett, JS</creator><creator>Tredway, TL</creator><creator>Dizikes, GJ</creator><creator>Nawrocki, JF</creator><general>Am Assoc Immnol</general><general>American Association of Immunologists</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T3</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>19940301</creationdate><title>Properties of a novel gene isolated from a Hodgkin's disease cell line that is expressed early during lymphoid cell activation</title><author>Bennett, JS ; Tredway, TL ; Dizikes, GJ ; Nawrocki, JF</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c464t-5463132824db1970d140a6c19d2f3284d93bc73d6a86c65f8e1138e96b20c5673</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>551000 -- Physiological Systems</topic><topic>activation</topic><topic>Amino Acid Sequence</topic><topic>ANIMAL CELLS</topic><topic>Base Sequence</topic><topic>BASIC BIOLOGICAL SCIENCES</topic><topic>Biological and medical sciences</topic><topic>BIOLOGICAL MATERIALS</topic><topic>BLOOD</topic><topic>BLOOD CELLS</topic><topic>BODY FLUIDS</topic><topic>cDNA</topic><topic>Cell Line, Transformed</topic><topic>cell lines</topic><topic>CLONING</topic><topic>CONNECTIVE TISSUE CELLS</topic><topic>DNA HYBRIDIZATION</topic><topic>DNA Primers - genetics</topic><topic>DNA SEQUENCING</topic><topic>DNA, Complementary - genetics</topic><topic>DNA, Neoplasm - genetics</topic><topic>DNA-CLONING</topic><topic>epag gene</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>GENE REGULATION</topic><topic>genes</topic><topic>Genes. Genome</topic><topic>Hodgkin Disease - genetics</topic><topic>Hodgkin Disease - immunology</topic><topic>Hodgkin's disease</topic><topic>Humans</topic><topic>HYBRIDIZATION</topic><topic>LEUKOCYTES</topic><topic>Lymphocyte Activation - genetics</topic><topic>LYMPHOCYTES</topic><topic>lymphocytes T</topic><topic>man</topic><topic>MATERIALS</topic><topic>METABOLIC ACTIVATION</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Mutation</topic><topic>nucleotide sequence</topic><topic>Open Reading Frames</topic><topic>prediction</topic><topic>SOMATIC CELLS</topic><topic>STRUCTURAL CHEMICAL ANALYSIS 550400 -- Genetics</topic><topic>T-Lymphocytes - immunology</topic><topic>Tumor Cells, Cultured - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bennett, JS</creatorcontrib><creatorcontrib>Tredway, TL</creatorcontrib><creatorcontrib>Dizikes, GJ</creatorcontrib><creatorcontrib>Nawrocki, JF</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Human Genome Abstracts</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>The Journal of immunology (1950)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bennett, JS</au><au>Tredway, TL</au><au>Dizikes, GJ</au><au>Nawrocki, JF</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Properties of a novel gene isolated from a Hodgkin's disease cell line that is expressed early during lymphoid cell activation</atitle><jtitle>The Journal of immunology (1950)</jtitle><addtitle>J Immunol</addtitle><date>1994-03-01</date><risdate>1994</risdate><volume>152</volume><issue>5</issue><spage>2229</spage><epage>2240</epage><pages>2229-2240</pages><issn>0022-1767</issn><eissn>1550-6606</eissn><coden>JOIMA3</coden><abstract>We have isolated a novel 667-bp cDNA clone, designated epag, from a Hodgkin's-disease cell line-derived library that is expressed in association with T cell activation and which is not related to any known gene family. By using reverse transcription/PCR, we have demonstrated that epag mRNA is expressed as early as 1 h after stimulation of normal PBMCs with anti-CD3. The levels of mRNA peaked by 4 h, and no expression was detectable by 12 h postactivation or in resting cells incubated in culture without activation. Expression of epag was also detected in PMA- and PHA-stimulated, but not in nonstimulated Jurkat cells, and overall its expression in transformed cell lines of hemopoietic origin is highly restricted. Sequence analysis of multiple independent cDNA clones showed that epag expressed in the Hodgkin's-disease cell line L428 is identical to the gene expressed in normal activated PBMC. Epag expression was detected by reverse transcription/PCR in RNA preparations made from various normal nonlymphoid tissues. Computer analysis of the sequence identified an open reading frame encoding a putative protein of 13.2 kDa initiating at a CUG translational codon. In vitro translation and Western blot analysis with anti-peptide serum supported this analysis. We hypothesize that epag functions as an early signal that helps mediate the activation of T cells.</abstract><cop>Bethesda, MD</cop><pub>Am Assoc Immnol</pub><pmid>8133036</pmid><doi>10.4049/jimmunol.152.5.2229</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record>
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ispartof The Journal of immunology (1950), 1994-03, Vol.152 (5), p.2229-2240
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subjects 551000 -- Physiological Systems
activation
Amino Acid Sequence
ANIMAL CELLS
Base Sequence
BASIC BIOLOGICAL SCIENCES
Biological and medical sciences
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
cDNA
Cell Line, Transformed
cell lines
CLONING
CONNECTIVE TISSUE CELLS
DNA HYBRIDIZATION
DNA Primers - genetics
DNA SEQUENCING
DNA, Complementary - genetics
DNA, Neoplasm - genetics
DNA-CLONING
epag gene
Fundamental and applied biological sciences. Psychology
Gene Expression
GENE REGULATION
genes
Genes. Genome
Hodgkin Disease - genetics
Hodgkin Disease - immunology
Hodgkin's disease
Humans
HYBRIDIZATION
LEUKOCYTES
Lymphocyte Activation - genetics
LYMPHOCYTES
lymphocytes T
man
MATERIALS
METABOLIC ACTIVATION
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Mutation
nucleotide sequence
Open Reading Frames
prediction
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS 550400 -- Genetics
T-Lymphocytes - immunology
Tumor Cells, Cultured - immunology
title Properties of a novel gene isolated from a Hodgkin's disease cell line that is expressed early during lymphoid cell activation
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