A novel DNMT3B splice variant expressed in tumor and pluripotent cells modulates genomic DNA methylation patterns and displays altered DNA binding

Suhasni Gopalakrishnan, Beth O. Van Emburgh, Jixiu Shan, Zhen Su, C. Robert Fields, Johannes Vieweg, Takashi Hamazaki, Philip H. Schwartz, Naohiro Terada, Keith D Robertson

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

DNA methylation is an epigenetic mark essential for mammalian development, genomic stability, and imprinting. DNA methylation patterns are established and maintained by three DNA methyltransferases: DNMT1, DNMT3A, and DNMT3B. Interestingly, all three DNMTs make use of alternative splicing. DNMT3B has nearly 40 known splice variants expressed in a tissue- and disease-specific manner, but very little is known about the role of these splice variants in modulatingDNMT3B function. We describe here the identification and characterization of a novel alternatively spliced form of DNMT3B lackingexon 5 within the NH 2-terminal regulatory domain. This variant, which we term DNMT3B3Δ5 because it is closely related in structure to the ubiquitously expressed DNMT3B3 isoform, is highly expressed in pluripotent cells and brain tissue, is downregulated during differentiation, and is conserved in the mouse. Creation of pluripotent iPS cells from fibroblasts results in marked induction of DNMT3B3Δ5. DNMT3B3Δ5 expression is also altered in human disease, with tumor cell lines displayingelevated or reduced expression depending on their tissue of origin. We then compared the DNA binding and subcellular localization of DNMT3B3Δ5 versus DNMT3B3, revealingthat DNMT3B3Δ5 possessed significantly enhanced DNA binding affinity and displayed an altered nuclear distribution. Finally, ectopic overexpression of DNMT3B3Δ5 resulted in repetitive element hypomethylation and enhanced cell growth in a colony formation assay. Taken together, these results show that DNMT3B3Δ5 may play an important role in stem cell maintenance or differentiation and suggest that sequences encoded by exon 5 influence the functional properties of DNMT3B.

Original languageEnglish (US)
Pages (from-to)1622-1634
Number of pages13
JournalMolecular Cancer Research
Volume7
Issue number10
DOIs
StatePublished - Oct 2009
Externally publishedYes

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DNA Methylation
DNA
Genomic Imprinting
Neoplasms
Genomic Instability
Alternative Splicing
Methyltransferases
Tumor Cell Line
Epigenomics
Exons
Protein Isoforms
Stem Cells
Down-Regulation
Fibroblasts
Maintenance
Brain
Growth

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Oncology

Cite this

A novel DNMT3B splice variant expressed in tumor and pluripotent cells modulates genomic DNA methylation patterns and displays altered DNA binding. / Gopalakrishnan, Suhasni; Van Emburgh, Beth O.; Shan, Jixiu; Su, Zhen; Fields, C. Robert; Vieweg, Johannes; Hamazaki, Takashi; Schwartz, Philip H.; Terada, Naohiro; Robertson, Keith D.

In: Molecular Cancer Research, Vol. 7, No. 10, 10.2009, p. 1622-1634.

Research output: Contribution to journalArticle

Gopalakrishnan, S, Van Emburgh, BO, Shan, J, Su, Z, Fields, CR, Vieweg, J, Hamazaki, T, Schwartz, PH, Terada, N & Robertson, KD 2009, 'A novel DNMT3B splice variant expressed in tumor and pluripotent cells modulates genomic DNA methylation patterns and displays altered DNA binding', Molecular Cancer Research, vol. 7, no. 10, pp. 1622-1634. https://doi.org/10.1158/1541-7786.MCR-09-0018
Gopalakrishnan, Suhasni ; Van Emburgh, Beth O. ; Shan, Jixiu ; Su, Zhen ; Fields, C. Robert ; Vieweg, Johannes ; Hamazaki, Takashi ; Schwartz, Philip H. ; Terada, Naohiro ; Robertson, Keith D. / A novel DNMT3B splice variant expressed in tumor and pluripotent cells modulates genomic DNA methylation patterns and displays altered DNA binding. In: Molecular Cancer Research. 2009 ; Vol. 7, No. 10. pp. 1622-1634.
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AU - Fields, C. Robert

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AU - Hamazaki, Takashi

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AU - Terada, Naohiro

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