PMID- 12616525
OWN - NLM
STAT- MEDLINE
DCOM- 20031120
LR  - 20231213
IS  - 0730-2312 (Print)
IS  - 0730-2312 (Linking)
VI  - 88
IP  - 5
DP  - 2003 Apr 1
TI  - Biochemical fractionation reveals association of DNA methyltransferase (Dnmt) 3b 
      with Dnmt1 and that of Dnmt 3a with a histone H3 methyltransferase and Hdac1.
PG  - 855-64
AB  - De novo DNA methyltransferases, Dnmt3a and 3b, were purified by fractionation of 
      S-100 extract from mouse lymphosarcoma cells through several chromatographic 
      matrices followed by glycerol density gradient centrifugation. Dnmt3a was 
      separated from Dnmt3b and Dnmt1 in the first column, Q-Sepharose whereas Dnmt3b 
      co-purified with Dnmt1 after further fractionation through Mono-S and Mono-Q 
      columns and glycerol density gradient centrifugation. Following purification, the 
      majority of de novo DNA methyltransfearse activity was associated with 
      Dnmt3b/Dnmt1 fractions. By contrast, the fractions containing Dnmt3a alone 
      exhibited markedly reduced activity, which correlated with diminished expression 
      of this isoform in these cells. Histone deacetylase 1(Hdac1) cofractionated with 
      Dnmt3a throughout purification whereas Hdac1 was separated from Dnmt3b/Dnmt1 
      following chromatography on Mono-Q column. Dnmt3a purified through glycerol 
      gradient centrifugation was also associated with a histone H3 methyltransferase 
      (HMTase) activity whereas purified Dnmt3b/Dnmt1 was devoid of any HMTase 
      activity. The activity of this HMTase was abolished when lysine 9 of N-terminal 
      histone H3 peptide was replaced by leucine whereas mutation of lysine 4 to 
      leucine inhibited this activity only partially. This is the first report on the 
      identification of a few key co-repressors associated with endogenous Dnmt3a and 
      of a complex containing Dnmt3b and a minor form of Dnmt1 following extensive 
      biochemical fractionation.
CI  - Copyright 2003 Wiley-Liss, Inc.
FAU - Datta, Jharna
AU  - Datta J
AD  - Department of Molecular and Cellular Biochemistry, College of Medicine, The Ohio 
      State University, Columbus, Ohio 43210, USA.
FAU - Ghoshal, Kalpana
AU  - Ghoshal K
FAU - Sharma, Sudarshana M
AU  - Sharma SM
FAU - Tajima, Shoji
AU  - Tajima S
FAU - Jacob, Samson T
AU  - Jacob ST
LA  - eng
GR  - P30 CA016058/CA/NCI NIH HHS/United States
GR  - CA81024/CA/NCI NIH HHS/United States
GR  - ES10874/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Cell Biochem
JT  - Journal of cellular biochemistry
JID - 8205768
RN  - 0 (DNMT3A protein, human)
RN  - 0 (Dnmt3a protein, mouse)
RN  - EC 2.1.1.- (Histone Methyltransferases)
RN  - EC 2.1.1.- (Methyltransferases)
RN  - EC 2.1.1.- (Protein Methyltransferases)
RN  - EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferase 1)
RN  - EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferases)
RN  - EC 2.1.1.37 (DNA Methyltransferase 3A)
RN  - EC 2.1.1.37 (DNMT1 protein, human)
RN  - EC 2.1.1.37 (Dnmt1 protein, mouse)
RN  - EC 2.1.1.43 (Histone-Lysine N-Methyltransferase)
RN  - EC 3.5.1.98 (HDAC1 protein, human)
RN  - EC 3.5.1.98 (Histone Deacetylase 1)
RN  - EC 3.5.1.98 (Histone Deacetylases)
SB  - IM
MH  - Animals
MH  - Centrifugation, Density Gradient
MH  - Chemical Fractionation
MH  - Chromatography, Agarose
MH  - DNA (Cytosine-5-)-Methyltransferase 1
MH  - DNA (Cytosine-5-)-Methyltransferases/*chemistry/isolation & purification
MH  - DNA Methyltransferase 3A
MH  - Histone Deacetylase 1
MH  - Histone Deacetylases/*chemistry/isolation & purification
MH  - Histone Methyltransferases
MH  - *Histone-Lysine N-Methyltransferase
MH  - Lymphoma, Non-Hodgkin
MH  - Methyltransferases/analysis/*chemistry
MH  - Mice
MH  - Protein Methyltransferases
MH  - Tumor Cells, Cultured/chemistry
MH  - DNA Methyltransferase 3B
EDAT- 2003/03/05 04:00
MHDA- 2003/12/03 05:00
CRDT- 2003/03/05 04:00
PHST- 2003/03/05 04:00 [pubmed]
PHST- 2003/12/03 05:00 [medline]
PHST- 2003/03/05 04:00 [entrez]
AID - 10.1002/jcb.10457 [doi]
PST - ppublish
SO  - J Cell Biochem. 2003 Apr 1;88(5):855-64. doi: 10.1002/jcb.10457.