Identification of a 5-Methylcytosine Site that may Regulate C/EBPβ Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets
Bactericidal/permeability-increasing protein (BPI) plays an important role in innate immune defense in mammals. A previous study showed that BPI gene expression correlates to gram-negative bacteria resistance. However, this gene showed tissue-specific expression in piglets and strongly expressed onl...
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| description | Bactericidal/permeability-increasing protein (BPI) plays an important role in innate immune defense in mammals. A previous study showed that
BPI
gene expression correlates to gram-negative bacteria resistance. However, this gene showed tissue-specific expression in piglets and strongly expressed only in the digestive tract. To investigate the mechanisms governing the tissue-specificity, bisulfite sequencing PCR and next generation sequencing were used for high accuracy methylation quantitation of CpG islands of
BPI
gene upstream in 11 different tissues from weaned Yorkshire piglets. Additionally, qPCR was used to examine mRNA levels of
BPI
gene as well as transcription factor. We additionally analyzed transcriptional regulation by studying key 5-methylcytosine sites and transcription factors. Results showed that
BPI
mRNA levels significantly correlated with the overall methylation as well as methylation at mC-15 which was non-CpG site, no significant correlation could be found between the
BPI
and transcription factor mRNA levels, EMSA test showed that C/EBPβ could interact with
BPI
wild-type promoter DNA, but not methylated DNA. So we confirmed that methylation of mC-15 residue could inhibit the ability of C/EBPβ binding to the
BPI
promoter and affect the expression, and this mechanism probably plays a role in the tissue specificity of
BPI
gene expression in weaned piglets. |
| doi_str_mv | 10.1038/srep28506 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4919782</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1799557934</sourcerecordid><originalsourceid>FETCH-LOGICAL-c410t-ca1f16de2c2d065d93ffe938383a350c0c6a2a1e6a9dfed6927fdce75de267013</originalsourceid><addsrcrecordid>eNptkd9u0zAUxiMEYtPYBS-AfAlIYbYTJ_ENEi1lVNpEtT_XlrFPUk-JndkOok-xd9mD8Ey4aqk2CfvCR_5-5zuWvyx7S_AngovmLHgYacNw9SI7prhkOS0offmkPspOQ7jDaTHKS8JfZ0e0LoqGNfw4e1hqsNG0RslonEWuRRKx_BLietOrTXTBWEDXJgKKaxnRIDfoCrqpl-lmfraYrf48opmx2tgOSavRV4jgh23TjQlhgvx6BLX1R4vfo4cQ9lPiGtBstUTnkFBj0cp0PcTwJnvVyj7A6f48yW6_LW7m3_OLH-fL-ZeLXJUEx1xJ0pJKA1VU44ppXrQt8KJJWxYMK6wqSSWBSnLdgq44rVutoGappaoxKU6yzzvfcfo5QJJs9LIXozeD9BvhpBHPFWvWonO_RMkJrxuaDN7vDby7nyBEMZigoO-lBTcFQWrOGat5USb0ww5V3oUUV3sYQ7DYZigOGSb23dN3Hch_iSXg4w4ISbIdeHHnJm_TX_3H7S_HiKjm</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1799557934</pqid></control><display><type>article</type><title>Identification of a 5-Methylcytosine Site that may Regulate C/EBPβ Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets</title><source>MEDLINE</source><source>Nature Free</source><source>DOAJ Directory of Open Access Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>Free Full-Text Journals in Chemistry</source><source>Springer Nature OA Free Journals</source><creator>Sun, Li ; Wang, Jing ; Yin, Xuemei ; Sun, Shouyong ; Zi, Chen ; Zhu, Guoqiang ; Wu, Shenglong ; Bao, Wenbin</creator><creatorcontrib>Sun, Li ; Wang, Jing ; Yin, Xuemei ; Sun, Shouyong ; Zi, Chen ; Zhu, Guoqiang ; Wu, Shenglong ; Bao, Wenbin</creatorcontrib><description>Bactericidal/permeability-increasing protein (BPI) plays an important role in innate immune defense in mammals. A previous study showed that
BPI
gene expression correlates to gram-negative bacteria resistance. However, this gene showed tissue-specific expression in piglets and strongly expressed only in the digestive tract. To investigate the mechanisms governing the tissue-specificity, bisulfite sequencing PCR and next generation sequencing were used for high accuracy methylation quantitation of CpG islands of
BPI
gene upstream in 11 different tissues from weaned Yorkshire piglets. Additionally, qPCR was used to examine mRNA levels of
BPI
gene as well as transcription factor. We additionally analyzed transcriptional regulation by studying key 5-methylcytosine sites and transcription factors. Results showed that
BPI
mRNA levels significantly correlated with the overall methylation as well as methylation at mC-15 which was non-CpG site, no significant correlation could be found between the
BPI
and transcription factor mRNA levels, EMSA test showed that C/EBPβ could interact with
BPI
wild-type promoter DNA, but not methylated DNA. So we confirmed that methylation of mC-15 residue could inhibit the ability of C/EBPβ binding to the
BPI
promoter and affect the expression, and this mechanism probably plays a role in the tissue specificity of
BPI
gene expression in weaned piglets.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/srep28506</identifier><identifier>PMID: 27338589</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>38 ; 38/22 ; 38/77 ; 38/90 ; 42/47 ; 45/29 ; 5-Methylcytosine - chemistry ; 631/208/1348 ; 631/208/199 ; 631/337/176/1988 ; 631/337/572/2102 ; 82 ; 82/29 ; 82/80 ; Animals ; Antimicrobial Cationic Peptides - chemistry ; Antimicrobial Cationic Peptides - genetics ; Antimicrobial Cationic Peptides - metabolism ; Base Sequence ; Blood Proteins - chemistry ; Blood Proteins - genetics ; Blood Proteins - metabolism ; CCAAT-Enhancer-Binding Protein-beta - chemistry ; CCAAT-Enhancer-Binding Protein-beta - genetics ; CCAAT-Enhancer-Binding Protein-beta - metabolism ; CpG Islands ; DNA Methylation ; Electrophoretic Mobility Shift Assay ; Gene Expression Regulation ; High-Throughput Nucleotide Sequencing ; Humanities and Social Sciences ; multidisciplinary ; Promoter Regions, Genetic ; Protein Binding ; Real-Time Polymerase Chain Reaction ; RNA, Messenger - metabolism ; Science ; Science (multidisciplinary) ; Sequence Analysis, DNA ; Swine ; Transcription Factors - genetics ; Transcription Factors - metabolism</subject><ispartof>Scientific reports, 2016-06, Vol.6 (1), p.28506-28506, Article 28506</ispartof><rights>The Author(s) 2016</rights><rights>Copyright © 2016, Macmillan Publishers Limited 2016 Macmillan Publishers Limited</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c410t-ca1f16de2c2d065d93ffe938383a350c0c6a2a1e6a9dfed6927fdce75de267013</citedby><cites>FETCH-LOGICAL-c410t-ca1f16de2c2d065d93ffe938383a350c0c6a2a1e6a9dfed6927fdce75de267013</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4919782/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4919782/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,27901,27902,41096,42165,51551,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27338589$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sun, Li</creatorcontrib><creatorcontrib>Wang, Jing</creatorcontrib><creatorcontrib>Yin, Xuemei</creatorcontrib><creatorcontrib>Sun, Shouyong</creatorcontrib><creatorcontrib>Zi, Chen</creatorcontrib><creatorcontrib>Zhu, Guoqiang</creatorcontrib><creatorcontrib>Wu, Shenglong</creatorcontrib><creatorcontrib>Bao, Wenbin</creatorcontrib><title>Identification of a 5-Methylcytosine Site that may Regulate C/EBPβ Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Bactericidal/permeability-increasing protein (BPI) plays an important role in innate immune defense in mammals. A previous study showed that
BPI
gene expression correlates to gram-negative bacteria resistance. However, this gene showed tissue-specific expression in piglets and strongly expressed only in the digestive tract. To investigate the mechanisms governing the tissue-specificity, bisulfite sequencing PCR and next generation sequencing were used for high accuracy methylation quantitation of CpG islands of
BPI
gene upstream in 11 different tissues from weaned Yorkshire piglets. Additionally, qPCR was used to examine mRNA levels of
BPI
gene as well as transcription factor. We additionally analyzed transcriptional regulation by studying key 5-methylcytosine sites and transcription factors. Results showed that
BPI
mRNA levels significantly correlated with the overall methylation as well as methylation at mC-15 which was non-CpG site, no significant correlation could be found between the
BPI
and transcription factor mRNA levels, EMSA test showed that C/EBPβ could interact with
BPI
wild-type promoter DNA, but not methylated DNA. So we confirmed that methylation of mC-15 residue could inhibit the ability of C/EBPβ binding to the
BPI
promoter and affect the expression, and this mechanism probably plays a role in the tissue specificity of
BPI
gene expression in weaned piglets.</description><subject>38</subject><subject>38/22</subject><subject>38/77</subject><subject>38/90</subject><subject>42/47</subject><subject>45/29</subject><subject>5-Methylcytosine - chemistry</subject><subject>631/208/1348</subject><subject>631/208/199</subject><subject>631/337/176/1988</subject><subject>631/337/572/2102</subject><subject>82</subject><subject>82/29</subject><subject>82/80</subject><subject>Animals</subject><subject>Antimicrobial Cationic Peptides - chemistry</subject><subject>Antimicrobial Cationic Peptides - genetics</subject><subject>Antimicrobial Cationic Peptides - metabolism</subject><subject>Base Sequence</subject><subject>Blood Proteins - chemistry</subject><subject>Blood Proteins - genetics</subject><subject>Blood Proteins - metabolism</subject><subject>CCAAT-Enhancer-Binding Protein-beta - chemistry</subject><subject>CCAAT-Enhancer-Binding Protein-beta - genetics</subject><subject>CCAAT-Enhancer-Binding Protein-beta - metabolism</subject><subject>CpG Islands</subject><subject>DNA Methylation</subject><subject>Electrophoretic Mobility Shift Assay</subject><subject>Gene Expression Regulation</subject><subject>High-Throughput Nucleotide Sequencing</subject><subject>Humanities and Social Sciences</subject><subject>multidisciplinary</subject><subject>Promoter Regions, Genetic</subject><subject>Protein Binding</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>RNA, Messenger - metabolism</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>Sequence Analysis, DNA</subject><subject>Swine</subject><subject>Transcription Factors - genetics</subject><subject>Transcription Factors - metabolism</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><recordid>eNptkd9u0zAUxiMEYtPYBS-AfAlIYbYTJ_ENEi1lVNpEtT_XlrFPUk-JndkOok-xd9mD8Ey4aqk2CfvCR_5-5zuWvyx7S_AngovmLHgYacNw9SI7prhkOS0offmkPspOQ7jDaTHKS8JfZ0e0LoqGNfw4e1hqsNG0RslonEWuRRKx_BLietOrTXTBWEDXJgKKaxnRIDfoCrqpl-lmfraYrf48opmx2tgOSavRV4jgh23TjQlhgvx6BLX1R4vfo4cQ9lPiGtBstUTnkFBj0cp0PcTwJnvVyj7A6f48yW6_LW7m3_OLH-fL-ZeLXJUEx1xJ0pJKA1VU44ppXrQt8KJJWxYMK6wqSSWBSnLdgq44rVutoGappaoxKU6yzzvfcfo5QJJs9LIXozeD9BvhpBHPFWvWonO_RMkJrxuaDN7vDby7nyBEMZigoO-lBTcFQWrOGat5USb0ww5V3oUUV3sYQ7DYZigOGSb23dN3Hch_iSXg4w4ISbIdeHHnJm_TX_3H7S_HiKjm</recordid><startdate>20160624</startdate><enddate>20160624</enddate><creator>Sun, Li</creator><creator>Wang, Jing</creator><creator>Yin, Xuemei</creator><creator>Sun, Shouyong</creator><creator>Zi, Chen</creator><creator>Zhu, Guoqiang</creator><creator>Wu, Shenglong</creator><creator>Bao, Wenbin</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</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>7X8</scope><scope>5PM</scope></search><sort><creationdate>20160624</creationdate><title>Identification of a 5-Methylcytosine Site that may Regulate C/EBPβ Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets</title><author>Sun, Li ; Wang, Jing ; Yin, Xuemei ; Sun, Shouyong ; Zi, Chen ; Zhu, Guoqiang ; Wu, Shenglong ; Bao, Wenbin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c410t-ca1f16de2c2d065d93ffe938383a350c0c6a2a1e6a9dfed6927fdce75de267013</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>38</topic><topic>38/22</topic><topic>38/77</topic><topic>38/90</topic><topic>42/47</topic><topic>45/29</topic><topic>5-Methylcytosine - chemistry</topic><topic>631/208/1348</topic><topic>631/208/199</topic><topic>631/337/176/1988</topic><topic>631/337/572/2102</topic><topic>82</topic><topic>82/29</topic><topic>82/80</topic><topic>Animals</topic><topic>Antimicrobial Cationic Peptides - chemistry</topic><topic>Antimicrobial Cationic Peptides - genetics</topic><topic>Antimicrobial Cationic Peptides - metabolism</topic><topic>Base Sequence</topic><topic>Blood Proteins - chemistry</topic><topic>Blood Proteins - genetics</topic><topic>Blood Proteins - metabolism</topic><topic>CCAAT-Enhancer-Binding Protein-beta - chemistry</topic><topic>CCAAT-Enhancer-Binding Protein-beta - genetics</topic><topic>CCAAT-Enhancer-Binding Protein-beta - metabolism</topic><topic>CpG Islands</topic><topic>DNA Methylation</topic><topic>Electrophoretic Mobility Shift Assay</topic><topic>Gene Expression Regulation</topic><topic>High-Throughput Nucleotide Sequencing</topic><topic>Humanities and Social Sciences</topic><topic>multidisciplinary</topic><topic>Promoter Regions, Genetic</topic><topic>Protein Binding</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>RNA, Messenger - metabolism</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Sequence Analysis, DNA</topic><topic>Swine</topic><topic>Transcription Factors - genetics</topic><topic>Transcription Factors - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sun, Li</creatorcontrib><creatorcontrib>Wang, Jing</creatorcontrib><creatorcontrib>Yin, Xuemei</creatorcontrib><creatorcontrib>Sun, Shouyong</creatorcontrib><creatorcontrib>Zi, Chen</creatorcontrib><creatorcontrib>Zhu, Guoqiang</creatorcontrib><creatorcontrib>Wu, Shenglong</creatorcontrib><creatorcontrib>Bao, Wenbin</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sun, Li</au><au>Wang, Jing</au><au>Yin, Xuemei</au><au>Sun, Shouyong</au><au>Zi, Chen</au><au>Zhu, Guoqiang</au><au>Wu, Shenglong</au><au>Bao, Wenbin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of a 5-Methylcytosine Site that may Regulate C/EBPβ Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2016-06-24</date><risdate>2016</risdate><volume>6</volume><issue>1</issue><spage>28506</spage><epage>28506</epage><pages>28506-28506</pages><artnum>28506</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Bactericidal/permeability-increasing protein (BPI) plays an important role in innate immune defense in mammals. A previous study showed that
BPI
gene expression correlates to gram-negative bacteria resistance. However, this gene showed tissue-specific expression in piglets and strongly expressed only in the digestive tract. To investigate the mechanisms governing the tissue-specificity, bisulfite sequencing PCR and next generation sequencing were used for high accuracy methylation quantitation of CpG islands of
BPI
gene upstream in 11 different tissues from weaned Yorkshire piglets. Additionally, qPCR was used to examine mRNA levels of
BPI
gene as well as transcription factor. We additionally analyzed transcriptional regulation by studying key 5-methylcytosine sites and transcription factors. Results showed that
BPI
mRNA levels significantly correlated with the overall methylation as well as methylation at mC-15 which was non-CpG site, no significant correlation could be found between the
BPI
and transcription factor mRNA levels, EMSA test showed that C/EBPβ could interact with
BPI
wild-type promoter DNA, but not methylated DNA. So we confirmed that methylation of mC-15 residue could inhibit the ability of C/EBPβ binding to the
BPI
promoter and affect the expression, and this mechanism probably plays a role in the tissue specificity of
BPI
gene expression in weaned piglets.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>27338589</pmid><doi>10.1038/srep28506</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Nature Free; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry; Springer Nature OA Free Journals |
| subjects | 38 38/22 38/77 38/90 42/47 45/29 5-Methylcytosine - chemistry 631/208/1348 631/208/199 631/337/176/1988 631/337/572/2102 82 82/29 82/80 Animals Antimicrobial Cationic Peptides - chemistry Antimicrobial Cationic Peptides - genetics Antimicrobial Cationic Peptides - metabolism Base Sequence Blood Proteins - chemistry Blood Proteins - genetics Blood Proteins - metabolism CCAAT-Enhancer-Binding Protein-beta - chemistry CCAAT-Enhancer-Binding Protein-beta - genetics CCAAT-Enhancer-Binding Protein-beta - metabolism CpG Islands DNA Methylation Electrophoretic Mobility Shift Assay Gene Expression Regulation High-Throughput Nucleotide Sequencing Humanities and Social Sciences multidisciplinary Promoter Regions, Genetic Protein Binding Real-Time Polymerase Chain Reaction RNA, Messenger - metabolism Science Science (multidisciplinary) Sequence Analysis, DNA Swine Transcription Factors - genetics Transcription Factors - metabolism |
| title | Identification of a 5-Methylcytosine Site that may Regulate C/EBPβ Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets |
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