Drosophila MOF controls Checkpoint protein2 and regulates genomic stability during early embryogenesis

BMC Molecular Biology

Manika Pal-Bhadra¹  Utpal Bhadra²  Debabani Roy Chowdhury²  M Janaki Ramaiah¹  Arpita Sarkar¹  Sreerangam NCVL Pushpavalli¹ 

[1] Centre for Chemical Biology, Indian Institute of Chemical Technology, Hyderabad, 500607, India;Functional Genomics and Gene Silencing Group, Centre for Cellular and Molecular Biology, Hyderabad, 500007, India

关键词: Anaphase bridges;    Chk2;    Drosophila melanogaster;    Syncytial embryos;    Mitosis;    Mof;   

Others  :  1091342
DOI  :  10.1186/1471-2199-14-1

 received in 2012-08-16, accepted in 2013-01-18,  发布年份 2013

【 摘 要 】

Background

In Drosophila embryos, checkpoints maintain genome stability by delaying cell cycle progression that allows time for damage repair or to complete DNA synthesis. Drosophila MOF, a member of MYST histone acetyl transferase is an essential component of male X hyperactivation process. Until recently its involvement in G2/M cell cycle arrest and defects in ionizing radiation induced DNA damage pathways was not well established.

Results

Drosophila MOF is highly expressed during early embryogenesis. In the present study we show that haplo-insufficiency of maternal MOF leads to spontaneous mitotic defects like mitotic asynchrony, mitotic catastrophe and chromatid bridges in the syncytial embryos. Such abnormal nuclei are eliminated and digested in the yolk tissues by nuclear fall out mechanism. MOF negatively regulates Drosophila checkpoint kinase 2 tumor suppressor homologue. In response to DNA damage the checkpoint gene Chk2 (Drosophila mnk) is activated in the mof mutants, there by causing centrosomal inactivation suggesting its role in response to genotoxic stress. A drastic decrease in the fall out nuclei in the syncytial embryos derived from mof¹/+; mnk^p6/+ females further confirms the role of DNA damage response gene Chk2 to ensure the removal of abnormal nuclei from the embryonic precursor pool and maintain genome stability. The fact that mof mutants undergo DNA damage has been further elucidated by the increased number of single and double stranded DNA breaks.

Conclusion

mof mutants exhibited genomic instability as evidenced by the occurance of frequent mitotic bridges in anaphase, asynchronous nuclear divisions, disruption of cytoskeleton, inactivation of centrosomes finally leading to DNA damage. Our findings are consistent to what has been reported earlier in mammals that; reduced levels of MOF resulted in increased genomic instability while total loss resulted in lethality. The study can be further extended using Drosophila as model system and carry out the interaction of MOF with the known components of the DNA damage pathway.

【 授权许可】

   
2013 Pushpavalli et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

Figure 9.

【 参考文献 】

• [1]Zhou BB, Elledge SJ: The DNA damage response: putting checkpoints in perspective. Nature 2000, 408:433-443.
• [2]Jaklevic BR, Su TT: Relative contribution of DNA repair, cell cycle check points and cell death to survival after DNA damage in Drosophila larvae. Curr Biol 2004, 14:23-32.
• [3]Voss AK, Thomas T: MYST family histone acetyltransferases take center stage in stem cells and development. Bioessays 2009, 31:1050-1061.
• [4]Morales V, Straub T, Neumann MF, Mengus G, Akhtar A, Becker PB: Functional integration of the histone acetyltransferase MOF into the dosage compensation complex. EMBO J 2004, 23:2258-2268.
• [5]Yang XJ: The diverse superfamily of lysine acetyltransferases and their roles in leukemia and other diseases. Nucleic Acids Res 2004, 32:959-976.
• [6]Smith ER, Pannuti A, Gu W, Steurnagel A, Cook RG, Allis CD, Lucchesi JC: The Drosophila MSL complex acetylates histone H4 at lysine 16, a chromatin modification linked to dosage compensation. Mol Cell Biol 2000, 20:312-318.
• [7]Smith ER, Cayrou C, Huang R, Lane WS, Cote J, Lucchesi JC: A human protein complex homologous to the Drosophila MSL complex is responsible for the majority of histone H4 acetylation at lysine 16. Mol Cell Biol 2005, 25:9175-9188.
• [8]Li X, Wu L, Corsa CA, Kunkel S, Dou Y: Two mammalian MOF complexes regulate transcription activation by distinct mechanisms. Mol Cell 2009, 36:290-301.
• [9]Hilfiker A, Hilfiker-Kleiner D, Pannuti A, Lucchesi JC: Mof, a putative acetyl transferase gene related to the Tip60 and MOZ human genes and to the SAS genes of yeast, is required for dosage compensation in Drosophila. EMBO J 1997, 16:2054-2060.
• [10]Taipale M, Rea S, Richter K, Vilar A, Lichter P, Imhof A, Akhtar A: hMOF histone acetyltransferase is required for histone H4 lysine 16 acetylation in mammalian cells. Mol Cell Biol 2005, 25:6798-6810.
• [11]Pal- Bhadra M, Horikoshi N, Pushpavalli Sreerangam NCVL, Sarkar A, Bag I, Krishnan A, Lucchesi JC, Kumar R, Yang Q, Pandita RJ, Singh M, Bhadra U, Eissenberg JC, Pandita TK: The role of MOF in the ionizing radiation response is conserved in Drosophila melanogaster. Chromosoma 2011. http://dx.doi.org/10.1007/s00412-011-0344-7 webcite
• [12]Shogren-Knaak M, Ishii H, Sun JM, Pazin MJ, Davie JR, Peterson CL: Histone H4-K16 acetylation controls chromatin structure and protein interactions. Science 2006, 311:844-847.
• [13]Corona DF, Clapier CR, Becker PB, Tamkun JW: Modulation of ISWI function by site-specific histoneacetylation. EMBO Rep 2002, 3:242-247.
• [14]Akhtar A, Becker PB: Activation of transcription through histone H4 acetylation by MOF, an acetyltransferase essential for dosage compensation in Drosophila. Mol Cell 2000, 5:367-375.
• [15]Gupta A, Guerin-Peyrou TG, Sharma GG, Park C, Agarwal M, Ganju RK, Pandita S, Choi K, Sukumar S, Pandita RK, Ludwig T, Pandita TK: The mammalian ortholog of Drosophila MOF that acetylates histone H4 lysine 16 is essential for embryogenesis and oncogenesis. Mol Cell Biol 2008, 28:397-409.
• [16]Fraga MF, Ballestar E, Villar-Garea A, Boix-Chornet M, Espada J, Schotta G, Bonaldi T, Haydon C, Ropero S, Petrie K, Iyer NG, Perez-Rosado A, Calvo E, Lopez JA, Cano A, Calasanz MJ, Colomer D, Piris MA, Ahn N, Imhof A, Caldas C, Jenuwein T, Esteller M: Loss of acetylation at Lys16 and trimethylation at Lys20 of histone H4 is a common hallmark of human cancer. Nat Genet 2005, 37:391-400.
• [17]Tram U, Riggs B, Sullivan W: Cleavage and gastrulation in Drosophila embryos. In Encyclopedia of Life Sciences. London: Macmillan Reference Ltd; 2002.
• [18]Foe VE, Odell GM Edgar BA, Bate M, Martinez Arias A: Mitosis and morphogenesis in the Drosophila embryo, Point and counterpoint. In The Development of Drosophila melanogaster vol 1. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press; 1993:149-300.
• [19]Dor E, Beck SA, Brock HW: Polycomb group mutants exhibit mitotic defects in syncytial cell cycles of Drosophila embryos. Dev Biol 2006, 29:312-322.
• [20]Fogarty P, Campbell SD, Abu-Shumays R, Phalle BS, Yu KR, Uy GL, Goldberg ML, Sullivan W: The Drosophila grapes gene is related to checkpoint gene chk1/rad27 and is required for late syncytial division fidelity. Curr Biol 1997, 7:418-426.
• [21]Sibon OC, Kelkar A, Lemstra W, Theurkauf WE: DNA replication/DNA-damage-dependent centrosome inactivation in Drosophila embryos. Nat Cell Biol 2000, 2:90-95.
• [22]Sibon OC, Stevenson VA, Theurkauf WE: DNA-replication checkpoint control at the Drosophila midblastula transition. Nature 1997, 388:93-97.
• [23]Sibon OC, Laurencon A, Hawley R, Theurkauf WE: The Drosophila ATM homologue Mei-41 has an essential checkpoint function at the midblastula transition. Curr Biol 1999, 9:302-312.
• [24]Takada S, Kelkar A, Theurkauf WE: Drosophila checkpoint kinase 2 couples centrosome function and spindle assembly to genomic integrity. Cell 2003, 113:87-99.
• [25]Oishi I, Sugiyama S, Otani H, Yamamura H, Nishida Y, Minami Y: A novel Drosophila nuclear protein serine/threonine kinase expressed in the germline during its establishment. Mech Dev 1998, 71:49-63.
• [26]Xu J, Xin S, Du W: Drosophila Chk2 is required for DNA damage-mediated cell cycle arrest and apoptosis. FEBS Lett 2001, 508:394-398.
• [27]Callaini G, Dallai R, Riparbelli MG: Cytochalasin induces spindle fusion in the syncytial blastoderm of the early Drosophila embryo. Biol Cell 1992, 74:249-254.
• [28]Nakayama M, Yamaguchi S, Sagisu Y, Sakurai H, Ito F, Kawasaki K: Loss of RecQ5 leads to spontaneous mitotic defects and chromosomal aberrations in Drosophila melanogaster. DNA Repair 2009, 8:232-241.
• [29]Sakurai H, Okado M, Fumiaki I, Kawasaki K: Anaphase DNA bridges induced by lack of RecQ5 in Drosophila syncytial embryos. FEBS Lett 2011, 585:1923-1928.
• [30]Sullivan W, Daily DR, Fogarty P, Yook KJ, Pimpinelli S: Delays in anaphase initiation occur in individual nuclei of the syncytial Drosophila embryo. Mol Biol Cell 1993, 4:885-896.
• [31]Roninson IB, Broude EV, Chang BD: If not apoptosis, then what? Treatment-induced senescence and mitotic catastrophe in tumor cells. Drug Resist Update 2001, 4:303-313.
• [32]Polo SE, Jackson SP: Dynamics of DNA damage response proteins at DNA breaks: a focus on protein modifications. Genes Dev 2011, 25:409-433.
• [33]Raff JW, Glover DM: Centrosomes, and not nuclei, initiate pole cell formation in Drosophila embryos. Cell 1989, 57:611-619.
• [34]Misteli T, Soutoglou E: The emerging role of nuclear architecture in DNA repair and genome maintenance. Nat Rev Mol Cell Biol 2009, 10:243-254.
• [35]Shi L, Oberdoerffer P: Chromatin dynamics in DNA double-strand break repair. Biochim Biophys Acta 2012, 1819:811-819.
• [36]Bakkenist CJ, Kastan MB: DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 2003, 421:499-506.
• [37]Bencokova Z, Kaufmann MR, Pires IM, Lecane PS, Giaccia AJ, Hammond EM: ATM activation and signaling under hypoxic conditions. Mol Cell Biol 2009, 29:526-537.
• [38]Hunt CR, Pandita RK, Laszlo A, Higashikubo R, Agarwal M, Kitamura T, Gupta A, Rie N, Horikoshi N, Baskaran R, Lee JH, Lobrich M, Paull TT, Roti JL: Hyperthermia activates a subset of ataxia-telangiectasia mutated effectors independent of DNA strand breaks and heat shock protein 70 status. Cancer Res 2007, 67:3010-3017.
• [39]Li J, Stern DF: DNA damage regulates Chk2 association with chromatin. J Biol Chem 2005, 280:37948-37956.
• [40]Takai H, Naka K, Okada Y, Watanabe M, Harada N, Saito S, Anderson CW, Appella E, Nakanishi M, Suzuki H, Nagashima K, Sawa H, Ikeda K, Motoyama N: Chk2-deficient mice exhibit radioresistance and defective p53-mediated transcription. EMBO J 2002, 21:5195-5205.
• [41]Chen LD, Gilkes M, Pan Y, Lane WS, Chen J: ATM and Chk2-dependent phosphorylation of MDMX contribute to p53 activation after DNA damage. EMBO J 2005, 24:3411-3422.