A VPE-like protease NtTPE8 exclusively expresses in the integumentary tapetum and is involved in seed development
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摘要
Programmed cell death(PCD) is an essential process for development, and shows conserved cytological features in both plants and animals. Caspases are well-known critical components of the PCD machinery in animals. However, currently few typical counterparts have been identified in plants and only several caspase-like proteases are known to be involved in plant PCD, indicating the existence of great challenge for confirming new caspase-like proteases and elucidating the mechanisms regulating plant PCD. Here, we report a novel cysteine protease, NtTPE8, which was extracted from tobacco seeds and confirmed as a new caspase-like protease.Recombinant NtTPE8 exhibited legumain and caspase-like proteolytic activities, both of which could be inhibited by the pan-caspase inhibitor(Z-VAD-FMK). Notably, NtTPE8 possessed several caspase activities and the capacity to cleave the cathepsin H substrate FVR, indicating a unique character of NtTPE8. NtTPE8 was exclusively expressed in the integumentary tapetum and thus, is the first specific molecular marker reported to date for this cell type. Downregulation of NtTPE8 caused seed abortion, via disturbing early embryogenesis, indicating its critical role in embryogenesis and seed development. In conclusion, we identified a novel caspase-like cysteine protease, NtTPE8,exclusively expressed in the integumentary tapetum that is involved in seed development.
Programmed cell death(PCD) is an essential process for development, and shows conserved cytological features in both plants and animals. Caspases are well-known critical components of the PCD machinery in animals. However, currently few typical counterparts have been identified in plants and only several caspase-like proteases are known to be involved in plant PCD, indicating the existence of great challenge for confirming new caspase-like proteases and elucidating the mechanisms regulating plant PCD. Here, we report a novel cysteine protease, NtTPE8, which was extracted from tobacco seeds and confirmed as a new caspase-like protease.Recombinant NtTPE8 exhibited legumain and caspase-like proteolytic activities, both of which could be inhibited by the pan-caspase inhibitor(Z-VAD-FMK). Notably, NtTPE8 possessed several caspase activities and the capacity to cleave the cathepsin H substrate FVR, indicating a unique character of NtTPE8. NtTPE8 was exclusively expressed in the integumentary tapetum and thus, is the first specific molecular marker reported to date for this cell type. Downregulation of NtTPE8 caused seed abortion, via disturbing early embryogenesis, indicating its critical role in embryogenesis and seed development. In conclusion, we identified a novel caspase-like cysteine protease, NtTPE8,exclusively expressed in the integumentary tapetum that is involved in seed development.
Programmed cell death(PCD) is an essential process for development, and shows conserved cytological features in both plants and animals. Caspases are well-known critical components of the PCD machinery in animals. However, currently few typical counterparts have been identified in plants and only several caspase-like proteases are known to be involved in plant PCD, indicating the existence of great challenge for confirming new caspase-like proteases and elucidating the mechanisms regulating plant PCD. Here, we report a novel cysteine protease, NtTPE8, which was extracted from tobacco seeds and confirmed as a new caspase-like protease.Recombinant NtTPE8 exhibited legumain and caspase-like proteolytic activities, both of which could be inhibited by the pan-caspase inhibitor(Z-VAD-FMK). Notably, NtTPE8 possessed several caspase activities and the capacity to cleave the cathepsin H substrate FVR, indicating a unique character of NtTPE8. NtTPE8 was exclusively expressed in the integumentary tapetum and thus, is the first specific molecular marker reported to date for this cell type. Downregulation of NtTPE8 caused seed abortion, via disturbing early embryogenesis, indicating its critical role in embryogenesis and seed development. In conclusion, we identified a novel caspase-like cysteine protease, NtTPE8,exclusively expressed in the integumentary tapetum that is involved in seed development.
引文
Belenghi B,Salomon M,Levine A(2004)Caspase-like activity in the seedlings of Pisum sativum eliminates weaker shoots during early vegetative development by induction of cell death.J Exp Bot 55:889-897
Bosch M,Franklin-Tong VE(2007)Temporal and spatial activation of caspase-like enzymes induced by selfincompatibility in Papaver pollen.Proc Natl Acad Sci USA 104:18327-18332
Bosch M,Poulter NS,Perry RM,Wilkins KA,Franklin-Tong VE(2010)Characterization of a legumain/vacuolar processing enzyme and YVADase activity in Papaver pollen.Plant Mol Biol 74:381-393
Bosch M,Poulter NS,Vatovec S,Franklin-Tong VE(2008)Initiation of programmed cell death in self-incompatibility:Role for cytoskeleton modifications and several caspaselike activities.Mol Plant 1:879-887
Bozhkov PV,Filonova LH,Suarez MF,Helmersson A,Smertenko AP,Zhivotovsky B,von Arnold S(2004)VEIDase is a principal caspase-like activity involved in plant programmed cell death and essential for embryonic pattern formation.Cell Death Differ 11:175-182
Chichkova NV,Kim SH,Titova ES,Kalkum M,Morozov VS,Rubtsov YP,Kalinina NO,Taliansky ME,Vartapetian AB(2004)A plant caspase-like protease activated during the hypersensitive response.Plant Cell 16:157-171
Chichkova NV,Shaw J,Galiullina RA,Drury GE,Tuzhikov AI,Kim SH,Kalkum M,Hong TB,Gorshkova EN,Torrance L,Vartapetian AB,Taliansky M(2010)Phytaspase,a relocalisable cell death promoting plant protease with caspase specificity.EMBO J 29:1149-1161
Coffeen WC,Wolpert TJ(2004)Purification and characterization of serine proteases that exhibit caspase-like activity and are associated with programmed cell death in Avena sativa.Plant Cell 16:857-873
Danon A,Rotari VI,Gordon A,Mailhac N,Gallois P(2004)Ultraviolet-C overexposure induces programmed cell death in Arabidopsis,which is mediated by caspase-like activities and which can be suppressed by caspase inhibitors,p35 and defender against apoptotic death.J Biol Chem 279:779-787
Ge Y,Cai YM,Bonneau L,Rotari V,Danon A,McKenzie EA,McLellan H,Mach L,Gallois P(2016)Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis.Cell Death Differ 23:1493-1501
Goldberg RB,Beals TP,Sanders PM(1993)Anther development:Basic principles and practical applications.Plant Cell5:1217-1229
Han JJ,Lin W,Oda Y,Cui KM,Fukuda H,He XQ(2012)The proteasome is responsible for caspase-3-like activity during xylem development.Plant J 72:129-141
Hatsugai N,Iwasaki S,Tamura K,Kondo M,Fuji K,Ogasawara K,Nishimura M,Hara-Nishimura I(2009)A novel membrane fusion-mediated plant immunity against bacterial pathogens.Gene Dev 23:2496-2506
Hatsugai N,Kuroyanagi M,Yamada K,Meshi T,Tsuda S,Kondo M,Nishimura M,Hara-Nishimura I(2004)A plant vacuolar protease,VPE,mediates virus-induced hypersensitive cell death.Science 305:855-858
He X,Kermode AR(2003)Proteases associated with programmed cell death of megagametophyte cells after germination of white spruce(Picea glauca)seeds.Plant Mol Biol 52:729-744
He X,Kermode AR(2010)Programmed cell death of the megagametophyte during post-germinative growth of white spruce(Picea glauca)seeds is regulated by reactive oxygen species and the ubiquitin-mediated proteolytic system.Plant Cell Physiol 51:1707-1720
Hiraiwa N,Nishimura M,Hara-Nishimura I(1999)Vacuolar processing enzyme is self-catalytically activated by sequential removal of the C-terminal and N-terminal propeptides.FEBS Lett 447:213-216
Hoeberichts FA,Woltering EJ(2003)Multiple mediators of plant programmed cell death:Interplay of conserved cell death mechanisms and plant-specific regulators.BioEssays 25:47-57
Ingram GC(2010)Family life at close quarters:Communication and constraint in angiosperm seed development.Protoplasma 247:195-214
Kapil RN,Tiwari SC(1978)The integumentary tapetum.Bot Rev 44:457-490
Korthout HA,Berecki G,Bruin W,van Duijn B,Wang M(2000)The presence and subcellular localization of caspase 3-like proteinases in plant cells.FEBS Lett 475:139-144
Kuroyanagi M,Nishimura M,Hara-Nishimura I(2002)Activation of Arabidopsis vacuolar processing enzyme by selfcatalytic removal of an auto-inhibitory domain of the C-terminal propeptide.Plant Cell Physiol 43:143-151
Lam E,del Pozo O(2000)Caspase-like protease involvement in the control of plant cell death.Plant Mol Biol 44:417-428
Li N,Zhang DS,Liu HS,Yin CS,Li XX,Liang WQ,Yuan Z,Xu B,Chu HW,Wang J,Wen TQ,Huang H,Luo D,Ma H,Zhang DB(2006)The rice tapetum degeneration retardation gene is required for tapetum degradation and anther development.Plant Cell 18:2999-3014
Linkies A,Graeber K,Knight C,Leubner-Metzger G(2010)The evolution of seeds.New Phytol 186:817-831
Liu JX,Howell SH(2010)Endoplasmic reticulum protein quality control and its relationship to environmental stress responses in plants.Plant Cell 22:2930-2942
Lopez-Fernandez MP,Maldonado S(2015)Programmed cell death in seeds of angiosperms.J Integr Plant Biol 57:996-1002
Lord CE,Gunawardena AH(2012)Programmed cell death in C.elegans,mammals and plants.Eur J Cell Biol 91:603-613
Nakaune S,Yamada K,Kondo M,Kato T,Tabata S,Nishimura M,Hara-Nishimura I(2005)A vacuolar processing enzyme,deltaVPE,is involved in seed coat formation at the early stage of seed development.Plant Cell 17:876-887
Piszczek E,Gutman W(2007)Caspase-like proteases and their role in programmed cell death in plants.Acta Physiol Plant29:391-398
Rojo E,Martin R,Carter C,Zouhar J,Pan S,Plotnikova J,Jin H,Paneque M,Sanchez-Serrano JJ,Baker B,Ausubel FM,Raikhel NV(2004)VPEgamma exhibits a caspase-like activity that contributes to defense against pathogens.Curr Biol 14:1897-1906
van Doorn WG,Beers EP,Dangl JL,Franklin-Tong VE,Gallois P,Hara-Nishimura I,Jones AM,Kawai-Yamada M,Lam E,Mundy J,Mur LA,Petersen M,Smertenko A,Taliansky M,Van Breusegem F,Wolpert T,Woltering E,Zhivotovsky B,Bozhkov PV(2011)Morphological classification of plant cell deaths.Cell Death Differ 18:1241-1246
Wang W,Zhao P,Zhou XM,Xiong HX,Sun MX(2015)Genomewide identification and characterization of cystatin family genes in rice(Oryza sativa L.).Plant Cell Rep 34:1579-1592
Wang W,Zhou XM,Xiong HX,Mao WY,Zhao P,Sun MX(2018)Papain-like and legumain-like proteases in rice:Genomewide identification,comprehensive gene feature characterization and expression analysis.BMC Plant Biol 18:87
Zhang D,Liu D,Lv X,Wang Y,Xun Z,Liu Z,Li F,Lu H(2014)The cysteine protease CEP1,a key executor involved in tapetal programmed cell death,regulates pollen development in Arabidopsis.Plant Cell 26:2939-2961
Zhang LR,Xu QX,Xing D,Gao CJ,Xiong HW(2009)Real-time detection of caspase-3-like protease activation in vivo using fluorescence resonance energy transfer during plant programmed cell death induced by ultraviolet C overexposure.Plant Physiol 150:1773-1783
Zhao P,Zhou XM,Zhang LY,Wang W,Ma LG,Yang LB,Peng XB,Bozhkov PV,Sun MX(2013)A bipartite molecular module controls cell death activation in the basal cell lineage of plant embryos.PLoS Biol 11:e1001655
Zhu J,Lou Y,Xu XF,Yang ZN(2011)A genetic pathway for tapetum development and function in Arabidopsis.J Integr Plant Biol 53:892-900
Bosch M,Franklin-Tong VE(2007)Temporal and spatial activation of caspase-like enzymes induced by selfincompatibility in Papaver pollen.Proc Natl Acad Sci USA 104:18327-18332
Bosch M,Poulter NS,Perry RM,Wilkins KA,Franklin-Tong VE(2010)Characterization of a legumain/vacuolar processing enzyme and YVADase activity in Papaver pollen.Plant Mol Biol 74:381-393
Bosch M,Poulter NS,Vatovec S,Franklin-Tong VE(2008)Initiation of programmed cell death in self-incompatibility:Role for cytoskeleton modifications and several caspaselike activities.Mol Plant 1:879-887
Bozhkov PV,Filonova LH,Suarez MF,Helmersson A,Smertenko AP,Zhivotovsky B,von Arnold S(2004)VEIDase is a principal caspase-like activity involved in plant programmed cell death and essential for embryonic pattern formation.Cell Death Differ 11:175-182
Chichkova NV,Kim SH,Titova ES,Kalkum M,Morozov VS,Rubtsov YP,Kalinina NO,Taliansky ME,Vartapetian AB(2004)A plant caspase-like protease activated during the hypersensitive response.Plant Cell 16:157-171
Chichkova NV,Shaw J,Galiullina RA,Drury GE,Tuzhikov AI,Kim SH,Kalkum M,Hong TB,Gorshkova EN,Torrance L,Vartapetian AB,Taliansky M(2010)Phytaspase,a relocalisable cell death promoting plant protease with caspase specificity.EMBO J 29:1149-1161
Coffeen WC,Wolpert TJ(2004)Purification and characterization of serine proteases that exhibit caspase-like activity and are associated with programmed cell death in Avena sativa.Plant Cell 16:857-873
Danon A,Rotari VI,Gordon A,Mailhac N,Gallois P(2004)Ultraviolet-C overexposure induces programmed cell death in Arabidopsis,which is mediated by caspase-like activities and which can be suppressed by caspase inhibitors,p35 and defender against apoptotic death.J Biol Chem 279:779-787
Ge Y,Cai YM,Bonneau L,Rotari V,Danon A,McKenzie EA,McLellan H,Mach L,Gallois P(2016)Inhibition of cathepsin B by caspase-3 inhibitors blocks programmed cell death in Arabidopsis.Cell Death Differ 23:1493-1501
Goldberg RB,Beals TP,Sanders PM(1993)Anther development:Basic principles and practical applications.Plant Cell5:1217-1229
Han JJ,Lin W,Oda Y,Cui KM,Fukuda H,He XQ(2012)The proteasome is responsible for caspase-3-like activity during xylem development.Plant J 72:129-141
Hatsugai N,Iwasaki S,Tamura K,Kondo M,Fuji K,Ogasawara K,Nishimura M,Hara-Nishimura I(2009)A novel membrane fusion-mediated plant immunity against bacterial pathogens.Gene Dev 23:2496-2506
Hatsugai N,Kuroyanagi M,Yamada K,Meshi T,Tsuda S,Kondo M,Nishimura M,Hara-Nishimura I(2004)A plant vacuolar protease,VPE,mediates virus-induced hypersensitive cell death.Science 305:855-858
He X,Kermode AR(2003)Proteases associated with programmed cell death of megagametophyte cells after germination of white spruce(Picea glauca)seeds.Plant Mol Biol 52:729-744
He X,Kermode AR(2010)Programmed cell death of the megagametophyte during post-germinative growth of white spruce(Picea glauca)seeds is regulated by reactive oxygen species and the ubiquitin-mediated proteolytic system.Plant Cell Physiol 51:1707-1720
Hiraiwa N,Nishimura M,Hara-Nishimura I(1999)Vacuolar processing enzyme is self-catalytically activated by sequential removal of the C-terminal and N-terminal propeptides.FEBS Lett 447:213-216
Hoeberichts FA,Woltering EJ(2003)Multiple mediators of plant programmed cell death:Interplay of conserved cell death mechanisms and plant-specific regulators.BioEssays 25:47-57
Ingram GC(2010)Family life at close quarters:Communication and constraint in angiosperm seed development.Protoplasma 247:195-214
Kapil RN,Tiwari SC(1978)The integumentary tapetum.Bot Rev 44:457-490
Korthout HA,Berecki G,Bruin W,van Duijn B,Wang M(2000)The presence and subcellular localization of caspase 3-like proteinases in plant cells.FEBS Lett 475:139-144
Kuroyanagi M,Nishimura M,Hara-Nishimura I(2002)Activation of Arabidopsis vacuolar processing enzyme by selfcatalytic removal of an auto-inhibitory domain of the C-terminal propeptide.Plant Cell Physiol 43:143-151
Lam E,del Pozo O(2000)Caspase-like protease involvement in the control of plant cell death.Plant Mol Biol 44:417-428
Li N,Zhang DS,Liu HS,Yin CS,Li XX,Liang WQ,Yuan Z,Xu B,Chu HW,Wang J,Wen TQ,Huang H,Luo D,Ma H,Zhang DB(2006)The rice tapetum degeneration retardation gene is required for tapetum degradation and anther development.Plant Cell 18:2999-3014
Linkies A,Graeber K,Knight C,Leubner-Metzger G(2010)The evolution of seeds.New Phytol 186:817-831
Liu JX,Howell SH(2010)Endoplasmic reticulum protein quality control and its relationship to environmental stress responses in plants.Plant Cell 22:2930-2942
Lopez-Fernandez MP,Maldonado S(2015)Programmed cell death in seeds of angiosperms.J Integr Plant Biol 57:996-1002
Lord CE,Gunawardena AH(2012)Programmed cell death in C.elegans,mammals and plants.Eur J Cell Biol 91:603-613
Nakaune S,Yamada K,Kondo M,Kato T,Tabata S,Nishimura M,Hara-Nishimura I(2005)A vacuolar processing enzyme,deltaVPE,is involved in seed coat formation at the early stage of seed development.Plant Cell 17:876-887
Piszczek E,Gutman W(2007)Caspase-like proteases and their role in programmed cell death in plants.Acta Physiol Plant29:391-398
Rojo E,Martin R,Carter C,Zouhar J,Pan S,Plotnikova J,Jin H,Paneque M,Sanchez-Serrano JJ,Baker B,Ausubel FM,Raikhel NV(2004)VPEgamma exhibits a caspase-like activity that contributes to defense against pathogens.Curr Biol 14:1897-1906
van Doorn WG,Beers EP,Dangl JL,Franklin-Tong VE,Gallois P,Hara-Nishimura I,Jones AM,Kawai-Yamada M,Lam E,Mundy J,Mur LA,Petersen M,Smertenko A,Taliansky M,Van Breusegem F,Wolpert T,Woltering E,Zhivotovsky B,Bozhkov PV(2011)Morphological classification of plant cell deaths.Cell Death Differ 18:1241-1246
Wang W,Zhao P,Zhou XM,Xiong HX,Sun MX(2015)Genomewide identification and characterization of cystatin family genes in rice(Oryza sativa L.).Plant Cell Rep 34:1579-1592
Wang W,Zhou XM,Xiong HX,Mao WY,Zhao P,Sun MX(2018)Papain-like and legumain-like proteases in rice:Genomewide identification,comprehensive gene feature characterization and expression analysis.BMC Plant Biol 18:87
Zhang D,Liu D,Lv X,Wang Y,Xun Z,Liu Z,Li F,Lu H(2014)The cysteine protease CEP1,a key executor involved in tapetal programmed cell death,regulates pollen development in Arabidopsis.Plant Cell 26:2939-2961
Zhang LR,Xu QX,Xing D,Gao CJ,Xiong HW(2009)Real-time detection of caspase-3-like protease activation in vivo using fluorescence resonance energy transfer during plant programmed cell death induced by ultraviolet C overexposure.Plant Physiol 150:1773-1783
Zhao P,Zhou XM,Zhang LY,Wang W,Ma LG,Yang LB,Peng XB,Bozhkov PV,Sun MX(2013)A bipartite molecular module controls cell death activation in the basal cell lineage of plant embryos.PLoS Biol 11:e1001655
Zhu J,Lou Y,Xu XF,Yang ZN(2011)A genetic pathway for tapetum development and function in Arabidopsis.J Integr Plant Biol 53:892-900