DNA methyltransferase protein synthesis is reduced in CXXC finger protein 1 deficient embryonic stem cells

Jill S. Butler, Lakshmi R. Palam, Courtney M. Tate, Jeremy R. Sanford, Ronald Wek, David G. Skalnik

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

CXXC finger protein 1 (CFP1) binds to unmethylated CpG dinucleotides and is required for embryogenesis. CFP1 is also a component of the Setd1A and Setd1B histone H3K4 methyltransferase complexes. Murine embryonic stem (ES) cells lacking CFP1 fail to differentiate, and exhibit a 70% reduction in global genomic cytosine methylation and a 50% reduction in DNA methyltransferase (DNMT1) protein and activity. This study investigated the underlying mechanism for reduced DNMT1 expression in CFP1-deficient ES cells. DNMT1 transcript levels were significantly elevated in ES cells lacking CFP1, despite the observed reduction in DNMT1 protein levels. To address the posttranscriptional mechanisms by which CFP1 regulates DNMT1 protein activity, pulse/chase analyses were carried out, demonstrating a modest reduction in DNMT1 protein half-life in CFP1-deficient ES cells. Additionally, global protein synthesis was decreased in ES cells lacking CFP1, contributing to a reduction in the synthesis of DNMT1 protein. ES cells lacking CFP1 were found to contain elevated levels of phosphorylated eIF2, and an accompanying reduction in translation initiation as revealed by a lower level of polyribosomes. These results reveal a novel role for CFP1 in the regulation of translation initiation, and indicate that loss of CFP1 function leads to decreased DNMT1 protein synthesis and half-life.

Original languageEnglish
Pages (from-to)223-231
Number of pages9
JournalDNA and Cell Biology
Volume28
Issue number5
DOIs
StatePublished - May 2009

Fingerprint

Protein Methyltransferases
Embryonic Stem Cells
Fingers
DNA
Proteins
Half-Life

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

DNA methyltransferase protein synthesis is reduced in CXXC finger protein 1 deficient embryonic stem cells. / Butler, Jill S.; Palam, Lakshmi R.; Tate, Courtney M.; Sanford, Jeremy R.; Wek, Ronald; Skalnik, David G.

In: DNA and Cell Biology, Vol. 28, No. 5, 05.2009, p. 223-231.

Research output: Contribution to journalArticle

Butler, Jill S. ; Palam, Lakshmi R. ; Tate, Courtney M. ; Sanford, Jeremy R. ; Wek, Ronald ; Skalnik, David G. / DNA methyltransferase protein synthesis is reduced in CXXC finger protein 1 deficient embryonic stem cells. In: DNA and Cell Biology. 2009 ; Vol. 28, No. 5. pp. 223-231.
@article{a7348845b0634bb4a03ad04c18086b2c,
title = "DNA methyltransferase protein synthesis is reduced in CXXC finger protein 1 deficient embryonic stem cells",
abstract = "CXXC finger protein 1 (CFP1) binds to unmethylated CpG dinucleotides and is required for embryogenesis. CFP1 is also a component of the Setd1A and Setd1B histone H3K4 methyltransferase complexes. Murine embryonic stem (ES) cells lacking CFP1 fail to differentiate, and exhibit a 70{\%} reduction in global genomic cytosine methylation and a 50{\%} reduction in DNA methyltransferase (DNMT1) protein and activity. This study investigated the underlying mechanism for reduced DNMT1 expression in CFP1-deficient ES cells. DNMT1 transcript levels were significantly elevated in ES cells lacking CFP1, despite the observed reduction in DNMT1 protein levels. To address the posttranscriptional mechanisms by which CFP1 regulates DNMT1 protein activity, pulse/chase analyses were carried out, demonstrating a modest reduction in DNMT1 protein half-life in CFP1-deficient ES cells. Additionally, global protein synthesis was decreased in ES cells lacking CFP1, contributing to a reduction in the synthesis of DNMT1 protein. ES cells lacking CFP1 were found to contain elevated levels of phosphorylated eIF2, and an accompanying reduction in translation initiation as revealed by a lower level of polyribosomes. These results reveal a novel role for CFP1 in the regulation of translation initiation, and indicate that loss of CFP1 function leads to decreased DNMT1 protein synthesis and half-life.",
author = "Butler, {Jill S.} and Palam, {Lakshmi R.} and Tate, {Courtney M.} and Sanford, {Jeremy R.} and Ronald Wek and Skalnik, {David G.}",
year = "2009",
month = "5",
doi = "10.1089/dna.2009.0854",
language = "English",
volume = "28",
pages = "223--231",
journal = "DNA and Cell Biology",
issn = "1044-5498",
publisher = "Mary Ann Liebert Inc.",
number = "5",

}

TY - JOUR

T1 - DNA methyltransferase protein synthesis is reduced in CXXC finger protein 1 deficient embryonic stem cells

AU - Butler, Jill S.

AU - Palam, Lakshmi R.

AU - Tate, Courtney M.

AU - Sanford, Jeremy R.

AU - Wek, Ronald

AU - Skalnik, David G.

PY - 2009/5

Y1 - 2009/5

N2 - CXXC finger protein 1 (CFP1) binds to unmethylated CpG dinucleotides and is required for embryogenesis. CFP1 is also a component of the Setd1A and Setd1B histone H3K4 methyltransferase complexes. Murine embryonic stem (ES) cells lacking CFP1 fail to differentiate, and exhibit a 70% reduction in global genomic cytosine methylation and a 50% reduction in DNA methyltransferase (DNMT1) protein and activity. This study investigated the underlying mechanism for reduced DNMT1 expression in CFP1-deficient ES cells. DNMT1 transcript levels were significantly elevated in ES cells lacking CFP1, despite the observed reduction in DNMT1 protein levels. To address the posttranscriptional mechanisms by which CFP1 regulates DNMT1 protein activity, pulse/chase analyses were carried out, demonstrating a modest reduction in DNMT1 protein half-life in CFP1-deficient ES cells. Additionally, global protein synthesis was decreased in ES cells lacking CFP1, contributing to a reduction in the synthesis of DNMT1 protein. ES cells lacking CFP1 were found to contain elevated levels of phosphorylated eIF2, and an accompanying reduction in translation initiation as revealed by a lower level of polyribosomes. These results reveal a novel role for CFP1 in the regulation of translation initiation, and indicate that loss of CFP1 function leads to decreased DNMT1 protein synthesis and half-life.

AB - CXXC finger protein 1 (CFP1) binds to unmethylated CpG dinucleotides and is required for embryogenesis. CFP1 is also a component of the Setd1A and Setd1B histone H3K4 methyltransferase complexes. Murine embryonic stem (ES) cells lacking CFP1 fail to differentiate, and exhibit a 70% reduction in global genomic cytosine methylation and a 50% reduction in DNA methyltransferase (DNMT1) protein and activity. This study investigated the underlying mechanism for reduced DNMT1 expression in CFP1-deficient ES cells. DNMT1 transcript levels were significantly elevated in ES cells lacking CFP1, despite the observed reduction in DNMT1 protein levels. To address the posttranscriptional mechanisms by which CFP1 regulates DNMT1 protein activity, pulse/chase analyses were carried out, demonstrating a modest reduction in DNMT1 protein half-life in CFP1-deficient ES cells. Additionally, global protein synthesis was decreased in ES cells lacking CFP1, contributing to a reduction in the synthesis of DNMT1 protein. ES cells lacking CFP1 were found to contain elevated levels of phosphorylated eIF2, and an accompanying reduction in translation initiation as revealed by a lower level of polyribosomes. These results reveal a novel role for CFP1 in the regulation of translation initiation, and indicate that loss of CFP1 function leads to decreased DNMT1 protein synthesis and half-life.

UR - http://www.scopus.com/inward/record.url?scp=65549109782&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=65549109782&partnerID=8YFLogxK

U2 - 10.1089/dna.2009.0854

DO - 10.1089/dna.2009.0854

M3 - Article

C2 - 19388845

AN - SCOPUS:65549109782

VL - 28

SP - 223

EP - 231

JO - DNA and Cell Biology

JF - DNA and Cell Biology

SN - 1044-5498

IS - 5

ER -