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Cell cycle regulation of DNMT1 and cancer

Cell cycle regulation of DNMT1 and cancer. M. M. The Genome and Epigenome Must be Faithfully Replicated with Each Cell Cycle. M. M. M. M. DNMT1. M. M. M. M. M. M. M. M. M. M. G0/G1. S. G2. M. DNMT1 transcription. Enzymatic activity. Steady state mRNA.

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Cell cycle regulation of DNMT1 and cancer

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  1. Cell cycle regulation of DNMT1 and cancer

  2. M M The Genome and Epigenome Must be Faithfully Replicated with Each Cell Cycle M M M M DNMT1 M M M M M M M M M M G0/G1 S G2 M DNMT1 transcription Enzymatic activity Steady state mRNA

  3. Expression of DNMT1 mRNA is regulated with the cell cycle

  4. G T A G T T T T T A T humanutr(5089-5408) C A C T G T G T G G G T G G C A A G T G A G A C T T G A T G T A G T T N T T A T rattusutr(5072-5093)(5123-5211) A T G T T G T A A T A T T T C daniorarioutr(2342-2364) A T G T T G T A A T A T T T C T T C A A A T A A A T C T C T C C T A T A A A c hic k enutr(4936-4991) A T G T T G T A A T A T T T C T T C A A A mouseutr(5076-5106)(5194-5233) A T G T T G T A A T A T T T C T T C A A A T A A A T C T C T C C T A T A A A C C humanutr(5089-5408) A T G T T G T A A T A T T T C T T C A A The DNMT1 3’UTR Contains A Highly Conserved Element - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - daniorarioutr(2342-2364) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - c hic k enutr(4936-4991) - - - - - - - - - - - - - - - - - - G T G T A G T A C T T T G T G C A T T C T G mouseutr(5076-5106)(5194-5233) T G A T T T A G T G A T C A A A T T G T G C A G T A C T T T G T G C A T T C T G humanutr(5089-5408) - - - - - - - - - - - - - C T T T G T G C A T T C T G rattusutr(5072-5093)(5123-5211) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - daniorarioutr(2342-2364) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - c hic k enutr(4936-4991) A A T T T T A A - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - mouseutr(5076-5106)(5194-5233) G A T T T T A A A A G T T T T T T A T T A T G C A T T A T A T C A A A T C T A C humanutr(5089-5408) A A T T T T A A - - - T T T T T T A T T C T G T A T T C T A T C A G A T C T G C rattusutr(5072-5093)(5123-5211) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - G T T T T T A T daniorarioutr(2342-2364) - - - - - - - - - - - - - - - - - - - - - - A C T T T A T G T A G T T T T T A T c hic k enutr(4936-4991) - - - - - - - - - - - - - - - - - - - - A G A C T T G A T G T A G T T N T T A T mouseutr(5076-5106)(5194-5233) C A C T G T A T G A G T G G A A A T T A A G A C T T T A T A rattusutr(5072-5093)(5123-5211) daniorarioutr(2342-2364) c hic k enutr(4936-4991) mouseutr(5076-5106)(5194-5233) A A A A A A A A A A A A A A A A A A A A A A A A A A humanutr(5089-5408) rattusutr(5072-5093)(5123-5211)

  5. serum induced 20h 0h kDa 84 41 32 A Specific Protein (p40) Binds the Conserved Region of the DNMT1 3’UTR in Arrested Cells 3’UTR probes / competitors 3’ 5’ UTR 5090-5408 5’- 259 UTR 5349-5405 D 5090-5350 3’- 56 UTR D 5090-5248 3’- 158 UTR D serum induction (h) UTRD 5’ 259 UTRD 3’ 158 UTRD 3 ’56 36 competitor: 16 20 24 0 8 none UTR kDa 130 85 43 32

  6. a a a a a a a DNMT1 3’UTR destabilizes mRNA in arrested cells +serum arrested ‭0 .5 1 2 4‬ ‭0 .5 1 2 4‬ 56 nt 1 t1/2=105 min 0.1 t1/2=58 min 0.01 ‭‬ 0 100 200 300

  7. DNMT1 DNMT1 DNMT1 DNMT1 UTR UTR UTR UTR D D DNMT1 DNMT1 56 56 UTR 56 D The Highly Conserved Region of the 3’UTR is Required to Downregulate the DNMT1 mRNA in Growth Arrested Cells serum induced (20h) growth arrested D D DNMT1 UTR D 5085 0 DNMT1 DNMT1-UTR 56 D 5350 DNMT1 5408 GFP CMV GFP 30 polyA pAD-DNMT-1 25 20 growth arrested Normalized DNMT1 mRNA Kan 15 serum induced (20h) 10 5 0 DNMT1 DNMT1 DNMT1 UTR D

  8. Deregulated cell cycle expression of DNMT1 leads to cellular transformation NIH 3T3 h-DNMT1 h-DNMT1-D3’UTR

  9. The 3’UTR inhibits ectopic expression of DNMT1 at the G0 phase of the cell cycle Ectopic expression of DNMT1 induces entry into the S phase of the cell cycle

  10. DNMT1 Mitogenic signals DNA replication DNA replication DNA methylation Protection of the DNA methylation pattern Fig. 2

  11. Inhibition of DNMT1 by antisense oligonucleotides inhibits Y1 tumor growth in vivo

  12. Nancy Detich QiangLi Zhuang Nadia Cervoni Ian Weaver Michael Meany

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