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桑梅香 河北医科大学第四医院 / 河北省肿瘤医院

Polo 样激酶 Plk3 对 p73 转录活性的影响 及对其磷酸化位点的分析. 桑梅香 河北医科大学第四医院 / 河北省肿瘤医院. Kinase Domain. PBD1. PBD2. Cdc5. 705. Plk1. 603. Saccharomyces cerevisiae. Plo1. 683. 685. Plk2. Schizosaccharomyces pombe. Plc1. 649. 646. Plk3. Caenorhabditis elegans. 576. Polo. Plk4. 907.

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桑梅香 河北医科大学第四医院 / 河北省肿瘤医院

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  1. Polo样激酶Plk3对p73转录活性的影响 及对其磷酸化位点的分析 桑梅香 河北医科大学第四医院/河北省肿瘤医院

  2. Kinase Domain PBD1 PBD2 Cdc5 705 Plk1 603 Saccharomyces cerevisiae Plo1 683 685 Plk2 Schizosaccharomyces pombe Plc1 649 646 Plk3 Caenorhabditis elegans 576 Polo Plk4 907 Drosophila melanogaster 598 Plx1 Xenopus laevis Mammals (Homo sapiens) Schematic representation of Polo and Polo-like kinases (Plks)

  3. P S Functions of Polo-Box Domain and Kinase Domain of Plks Polo-Box Domain Kinase Domain PBD1 PBD2 PB2 Kinase PB1

  4. DNA Damage DNA Damage ATM(ATR) ATM(ATR) Plk1 Plk3 ? ? ? p53 p53 p63 p63 p73 p73 Polo-like kinases (Plks) and p53 family

  5. DAPY HA-p73 FLAG-Plk3 Merge Co-localization of Plk3 and p73 in cells

  6. １　　 54 62 p73α(1-62) １　　 54 130 p73α(1-130) TA DBD OD SAM 114 131 310 328 A p73α(114-328) 311 345 380 483 p73α(311-483) p73α(469-636) 469 484 549 636 GST-p73α GST-p73α 114-328 311-483 469-636 114-328 311-483 469-636 1-130 kDa GST 1-62 kDa 1-130 GST 1-62 75- 50- 〔35S〕FLAG-Plk3 37- 50- 25- Interaction between Plk3 and p73α B C

  7. HA-p73α - + + + + - FLAG-Plk3 - - HA-p73α - + + + + - FLAG-Plk3 - - HA-p73α - + + + + - FLAG-Plk3 - - HA-p73α - + + + FLAG-Plk3 - - p21WAF1 Bax p53AIP1 Plk3 p73α GAPDH Plk3 inhibits p73α-mediated transcription activation

  8. HA-p73α - + + + - Plk3(K52R)- - HA-p73α - + + + - Plk3(K52R)- - HA-p73α - + + + Plk3(K52R) - - p21WAF1 Bax HA-p73α - + + + - Plk3(K52R)- - p53AIP1 Plk3 p73α GAPDH Kinase-deficient Plk3 (K52R) fails to inhibit p73α-mediated transcription activation

  9. Plk3 inhibits p73α-mediated transcription activation H1299 (p53 null) pcDNA3 p73 p73/Plk3 p73/Plk3(K52R)

  10. HA-p73α+Plk3(K52R) HA-p73α+Plk3 Plk3(K52R) FLAG-Plk3 pcDNA3 HA-p73α PARP cleaved PARP ► Actin CDDP (10μM) Plk3 inhibits the p73α-mediated apoptosis

  11. TA DBD OD SAM １　　 54 62 p73α(1-62) １　　 54 130 p73α(1-130) 114 131 310 328 p73α(114-328) 311 345 380 483 p73α(311-483) 469 484 549 636 p73α(469-636) GST-p73α GST-p73α 114-328 311-483 469-636 kDa 1-130 GST 1-62 Control 311-483 469-636 114-328 1-130 GST 1-62 50- 37- 25- Plk3 phosphorylates p73 at its N-terminal region in vitro A B C

  12. amino residues of p73 between 63-113: AQFNLLSSTMDQMSSRAASASPYTPEHAASVPTHSPYAQPSSTFDTMS PAP This short fragment include 11 Ser residus and 5 Thr residues. A GST-p73α 1-130 (T86A) 311-483 469-636 Control 114-328 GST 1-62 T86 is not the phosphorylation sites of p73 targeted by Plk3

  13. Conclusions • Plk3 inhibits p73α-mediated transcription activation, whereas kinase-deficient Plk3 fails to inhibit p73α-mediated transcription activation. • Plk3 inhibits the p73α-mediated apoptosis through physical interaction and phosphorylation.

  14. Thank You For Your Attention!

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