DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts

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dc.contributor.authorPARKER, M Iqbal
dc.contributor.authorde Haa, Judy B
dc.contributor.authorGevers, Wieland
dc.date.accessioned2021-10-08T07:20:44Z
dc.date.available2021-10-08T07:20:44Z
dc.date.issued1986
dc.description.abstractSodium butyrate is very often used to alter gene expression in cultured cells. In this study, we examined the effects of this compound on various cellular events in WI-38 human embryonic lung fibroblasts in culture. During a 16-20-h treatment at sodium butyrate concentrations of between 5 and 20 mM, no adverse effects on cell morphology were observed. However, cell division and DNA synthesis were reversibly inhibited, the latter by 85, 80, and 70% at sodium butyrate concentrations of 5, 10, and 20 mM, respectively. Although overall protein synthetic activity was not significantly affected, RNA synthesis decreased to 76% of the control values at a sodium butyrate concentration of 5 mM. Butyrate treatment also caused hypermethylation of DNA cytosines as determined by differential digestion by MspI/HpaII restriction endonucleases and by high performance liquid chromatography analysis of the DNA. The 5-methylcytosine content of the DNA in untreated WI-38 fibroblasts was 2.94 +/- 0.46% of total cytosine residues, while in cultures treated with 5, 10, and 20 mM sodium butyrate, these values were 5.76 +/- 0.28, 5.91 +/- 0.37, and 6.8 +/- 0.44%, respectively. An interesting feature is that this hypermethylation occurred in DNA which was synthesized in the presence of sodium butyrate (newly synthesized) as well as in DNA which had been synthesized before butyrate administration (pre-existing DNA). The hypermethylated state was conserved only in the former situation, since the methylcytosines were rapidly lost in the subsequent generation in the latter case. It would therefore appear that methylcytosines are maintained after cell replication only if they are generated on newly synthesized DNA.
dc.identifier.apacitationPARKER, M. I., de Haa, J. B., & Gevers, W. (1986). DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts. <i>The Journal of Biological Chemistry</i>, 261(6), 2786 - 27890. http://hdl.handle.net/11427/34992en_ZA
dc.identifier.chicagocitationPARKER, M Iqbal, Judy B de Haa, and Wieland Gevers "DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts." <i>The Journal of Biological Chemistry</i> 261, 6. (1986): 2786 - 27890. http://hdl.handle.net/11427/34992en_ZA
dc.identifier.citationPARKER, M.I., de Haa, J.B. & Gevers, W. 1986. DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts. <i>The Journal of Biological Chemistry.</i> 261(6):2786 - 27890. http://hdl.handle.net/11427/34992en_ZA
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.ris TY - Journal Article AU - PARKER, M Iqbal AU - de Haa, Judy B AU - Gevers, Wieland AB - Sodium butyrate is very often used to alter gene expression in cultured cells. In this study, we examined the effects of this compound on various cellular events in WI-38 human embryonic lung fibroblasts in culture. During a 16-20-h treatment at sodium butyrate concentrations of between 5 and 20 mM, no adverse effects on cell morphology were observed. However, cell division and DNA synthesis were reversibly inhibited, the latter by 85, 80, and 70% at sodium butyrate concentrations of 5, 10, and 20 mM, respectively. Although overall protein synthetic activity was not significantly affected, RNA synthesis decreased to 76% of the control values at a sodium butyrate concentration of 5 mM. Butyrate treatment also caused hypermethylation of DNA cytosines as determined by differential digestion by MspI/HpaII restriction endonucleases and by high performance liquid chromatography analysis of the DNA. The 5-methylcytosine content of the DNA in untreated WI-38 fibroblasts was 2.94 +/- 0.46% of total cytosine residues, while in cultures treated with 5, 10, and 20 mM sodium butyrate, these values were 5.76 +/- 0.28, 5.91 +/- 0.37, and 6.8 +/- 0.44%, respectively. An interesting feature is that this hypermethylation occurred in DNA which was synthesized in the presence of sodium butyrate (newly synthesized) as well as in DNA which had been synthesized before butyrate administration (pre-existing DNA). The hypermethylated state was conserved only in the former situation, since the methylcytosines were rapidly lost in the subsequent generation in the latter case. It would therefore appear that methylcytosines are maintained after cell replication only if they are generated on newly synthesized DNA. DA - 1986 DB - OpenUCT DP - University of Cape Town IS - 6 J1 - The Journal of Biological Chemistry LK - https://open.uct.ac.za PY - 1986 SM - 0021-9258 SM - 1083-351X T1 - DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts TI - DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts UR - http://hdl.handle.net/11427/34992 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/34992
dc.identifier.vancouvercitationPARKER MI, de Haa JB, Gevers W. DNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts. The Journal of Biological Chemistry. 1986;261(6):2786 - 27890. http://hdl.handle.net/11427/34992.en_ZA
dc.language.isoeng
dc.publisher.departmentDepartment of Medicine
dc.publisher.facultyFaculty of Health Sciences
dc.sourceThe Journal of Biological Chemistry
dc.source.journalissue6
dc.source.journalvolume261
dc.source.pagination2786 - 27890
dc.source.urihttps://dx.doi.org/10.7196/sajs.718
dc.subject.otherButyrates
dc.subject.otherButyric Acid
dc.subject.otherCell Division
dc.subject.otherCell Line
dc.subject.otherChromatography, High Pressure Liquid
dc.subject.otherDNA (Cytosine-5-)-Methyltransferases
dc.subject.otherDNA Replication
dc.subject.otherFibroblasts
dc.subject.otherHumans
dc.subject.otherProtein Biosynthesis
dc.subject.otherRNA
dc.subject.otherUridine
dc.subject.otherButyrates
dc.subject.otherButyric Acid
dc.subject.otherRNA
dc.titleDNA hypermethylation in sodium butyrate-treated WI-38 fibroblasts
dc.typeJournal Article
uct.type.publicationResearch
uct.type.resourceJournal Article
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