玉米生物钟基因ZmPRR1-2互作蛋白质的筛选

doi:10.3969/j.issn.1004-1524.20240204

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (5): 977-986.DOI: 10.3969/j.issn.1004-1524.20240204

玉米生物钟基因ZmPRR1-2互作蛋白质的筛选

王闻琦(), 王盼盼, 张严玲, 刘青青, 洪双双, 赵高鹏, 刘泓畅, 王翠玲()

河南科技大学农学院,河南洛阳 471023

收稿日期:2024-03-04 出版日期:2025-05-25 发布日期:2025-06-11
作者简介:王闻琦(1998—),男,河南许昌人,硕士研究生,从事作物分子育种相关研究。E-mail:945987039@qq.com
通讯作者: *王翠玲,E-mail: lywgg@126.com
基金资助:
国家自然科学基金河南省联合基金(U2004153)

Screening of proteins interacting with circadian clock gene ZmPRR1-2 in maize

WANG Wenqi(), WANG Panpan, ZHANG Yanling, LIU Qingqing, HONG Shuangshuang, ZHAO Gaopeng, LIU Hongchang, WANG Cuiling()

School of Agriculture, Henan University of Science and Technology, Luoyang 471023, Henan, China

Received:2024-03-04 Online:2025-05-25 Published:2025-06-11

摘要/Abstract

摘要：

伪应答调节蛋白TOC1是伪应答调节因子(pseudo-response regulator, PRR)家族5个成员之一,作为生物钟中央振荡器的核心组成部分,与MYB类转录因子LHY和CCA1构成了中央振荡器的中心负反馈调控环,在生物钟系统中起着重要作用。ZmPRR1-2基因是拟南芥TOC1的同源基因,为了深入挖掘ZmPRR1-2的生物学功能,本研究利用酵母双杂交筛库技术,从前期构建的长日照诱导的热带玉米自交系的cDNA文库中筛选与ZmPRR1-2互作的蛋白质。结果表明,构建的诱饵载体pGBKT7-ZmPRR1-2对酵母菌株无毒性,且对报告基因无自激活活性。筛选鉴定出12个与ZmPRR1-2互作的蛋白质。这些候选互作蛋白质的功能涉及抗氧化反应、水缺乏响应、铝离子响应、光合作用、电子传递链、光收集、生长素激活信号通路、DNA模板转录的调控、色氨酸生物合成过程、翻译、免疫反应、凋亡信号通路的正向调节、参与蛋白质分解代谢过程的蛋白质水解等多个方面。推测ZmPRR1-2蛋白与以上蛋白质互作参与多个信号转导和代谢途径,该结果为进一步探索ZmPRR1-2基因的分子功能与调控网络奠定了基础。

关键词: 玉米, 生物钟, ZmPRR1-2, 酵母双杂交, 互作蛋白质

Abstract:

The pseudo-response regulatory protein TOC1, one of the five members of the pseudo-response regulator (PRR) family proteins, as the core component of the central oscillator of clock, together with the MYB-like transcription factors LHY and CCA1, constitute the central negative regulatory feedback loop of the central oscillator of clock, which plays an important role in the circadian clock system. The ZmPRR1-2 gene is a homologous gene of TOC1 in Arabidopsis. In order to deeply explore the biological function of ZmPRR1-2, the proteins interacting with ZmPRR1-2 was screened from the yeast cDNA library of a tropical maize inbred line induced by long-day photoperiod treatment by yeast two-hybrid technique. The results showed that the bait vector PGBKT7-ZmPRR1-2 was not toxic to yeast strain and had no self-activation effect on the reporter gene. A total of 12 proteins containing the full length interacting with ZmPRR1-2 were identified. These candidate interacting proteins were involved in many pathways, such as response to oxidative stress, response to oxidative stress, response to aluminum ion, photosynthesis, electron transport chain, light harvesting, auxin-activated signaling pathway, regulation of DNA-templated transcription, tryptophan biosynthetic process, translation, immune response, positive regulation of apoptotic signaling pathway and proteolysis involved in protein catabolic process. It is speculated that ZmPRR1-2 protein may participate in multiple signal transduction and metabolic pathways by interacting with the identified interaction proteins. The results might improve our understanding of the further study of the molecular function and regulatory mechanism of ZmPRR1-2.

Key words: maize, circadian, ZmPRR1-2, yeast two-hybrid system, interacting protein

中图分类号:

S513

王闻琦, 王盼盼, 张严玲, 刘青青, 洪双双, 赵高鹏, 刘泓畅, 王翠玲. 玉米生物钟基因ZmPRR1-2互作蛋白质的筛选[J]. 浙江农业学报, 2025, 37(5): 977-986.

WANG Wenqi, WANG Panpan, ZHANG Yanling, LIU Qingqing, HONG Shuangshuang, ZHAO Gaopeng, LIU Hongchang, WANG Cuiling. Screening of proteins interacting with circadian clock gene ZmPRR1-2 in maize[J]. Acta Agriculturae Zhejiangensis, 2025, 37(5): 977-986.

图/表 6

参考文献 30

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基因名称 Gene name	编码蛋白质 Coding protein	染色体号码 Chr.No.	CDS长度 CDS length/bp	氨基酸个数 Number of amino acid	生物进程 Biological process	分子功能 Molecular function	亚细胞定位 Subcellular location
ZmAASR2	脱落酸胁迫成熟2 Abscisic acid stress ripening2	2	396	132	转录调控,DNA 模板化 Regulation of transcription, DNA-templated 抗氧化反应 Response to oxidative stress 水缺乏响应 Response to water deprivation 铝离子响应 Response to aluminum ion	序列特异性 DNA结合 Sequence-specific DNA binding	细胞核 Nucleolus 细胞质 Cytoplasm
ZmPET7	光系统Ⅰ反应中心亚基Ⅱ Photosystem Ⅰ reaction center subunit Ⅱ	1	600	200	光合作用 Photosynthesis	未知 Unknown	叶绿体类囊体膜 Chloroplast thylakoid membrane
ZmUCC3	Uclacyanin-3	10	510	170	电子传递链 Electron transport chain	电子传递活性 Electron transfer activity DNA结合 DNA binding	膜 Membrane
ZmPSPB1	光系统Ⅱ出氧系统的23 ku 亚基 23 ku subunit of oxygen evolving system of photosystem Ⅱ	2	777	259	光合作用 Photosynthesis	钙离子结合 Calcium ion binding 2-烯醛还原酶 [NAD(P)+]活性 2-Alkenal reductase [NAD(P)+] activity	叶绿体 Chloroplast
ZmLHCB2	叶绿素a-b结合蛋白 Chlorophyll a-b binding protein	7	789	263	光合作用,光收集 Photosynthesis, light harvesting	叶绿素结合 Chlorophyll binding	叶绿体类囊体膜 Chloroplast thylakoid membrane
ZmRUM1	生长素响应蛋白 Auxin- responsive protein	3	363	121	生长素激活信号通路 Auxin-activated signaling pathway DNA 模板转录的调控 Regulation of DNA- templated transcription	未知 Unknown	细胞核 Nucleolus
ZmTSAH1	色氨酸合成酶 Tryptophan synthase	1	1 020	340	色氨酸生物合成过程 Tryptophan biosynthetic process	色氨酸合成酶活性 Tryptophan synthase activity 吲哚-3-甘油-磷酸裂解酶活性 Indole-3-glycerol- phosphate lyase activity	叶绿体 Chloroplast 胞液 Cytosol
ZmNH4	nudix 水解酶 Nudix hydrolase 4	9	435	145	未知 Unknown	无机焦磷酸酶活性 Pyrophosphatase activity 水解酶活性 Hydrolase activity 金属离子结合 Metal ion binding	细胞质 Cytoplasm 细胞核 Nucleolus
ZmCL187-21A	60S核糖体蛋白L13 60S ribosomal protein L13	3	627	209	翻译 Translation	RNA结合 RNA binding 核糖体的结构成分 Structural constituent of ribosome	细胞质大核糖体亚基 Cytosolic large ribosomal subunit
ZmCCP2	半胱氨酸蛋白酶 Cysteine proteinase 2	7	1 083	361	免疫反应 Immune response 凋亡信号通路的正向调节 Positive regulation of apoptotic signaling pathway 参与蛋白质分解代谢过程的蛋白质水解 Proteolysis involved in protein catabolic process	半胱氨酸型内肽酶活性 Cysteine-type endopeptidase activity 苯丙氨酸转氨酶活性 Lyase activity	细胞外间隙 Extracellular space 溶酶体 Lysosome
ZmCSU27	紫外线-B-抑制蛋白 Ultraviolet-B- repressible protein	7	369	123	光合作用 Photosynthesis	蛋白质结构域特异性结合 Protein domain specific binding	细胞膜 Cytomembrane 叶绿体类囊体膜 Chloroplast thylakoid membrane
ZmNDF4	电子载体/电子传递体/铁离子结合蛋白 Electron carrier/ electron transporter/ iron ion binding protein	7	576	192	电子传递链 Electron transport chain 含P450的电子传递链 P450-containing electron transport chain 光系统Ⅰ中的光合电子传递 Photosynthetic electron transport in photosystem Ⅰ	2个铁原子, 2个硫原子簇结合 2 Iron, 2 sulfur cluster binding 电子传递活性 Electron transfer activity 金属离子结合 Metal ion binding 铁硫团簇结合 Iron-sulfur cluster binding	叶绿体类囊体膜 Chloroplast thylakoid membrane 胞液 Cytosol

基因名称 Gene name	编码蛋白质 Coding protein	染色体号码 Chr.No.	CDS长度 CDS length/bp	氨基酸个数 Number of amino acid	生物进程 Biological process	分子功能 Molecular function	亚细胞定位 Subcellular location
ZmAASR2	脱落酸胁迫成熟2 Abscisic acid stress ripening2	2	396	132	转录调控,DNA 模板化 Regulation of transcription, DNA-templated 抗氧化反应 Response to oxidative stress 水缺乏响应 Response to water deprivation 铝离子响应 Response to aluminum ion	序列特异性 DNA结合 Sequence-specific DNA binding	细胞核 Nucleolus 细胞质 Cytoplasm
ZmPET7	光系统Ⅰ反应中心亚基Ⅱ Photosystem Ⅰ reaction center subunit Ⅱ	1	600	200	光合作用 Photosynthesis	未知 Unknown	叶绿体类囊体膜 Chloroplast thylakoid membrane
ZmUCC3	Uclacyanin-3	10	510	170	电子传递链 Electron transport chain	电子传递活性 Electron transfer activity DNA结合 DNA binding	膜 Membrane
ZmPSPB1	光系统Ⅱ出氧系统的23 ku 亚基 23 ku subunit of oxygen evolving system of photosystem Ⅱ	2	777	259	光合作用 Photosynthesis	钙离子结合 Calcium ion binding 2-烯醛还原酶 [NAD(P)+]活性 2-Alkenal reductase [NAD(P)+] activity	叶绿体 Chloroplast
ZmLHCB2	叶绿素a-b结合蛋白 Chlorophyll a-b binding protein	7	789	263	光合作用,光收集 Photosynthesis, light harvesting	叶绿素结合 Chlorophyll binding	叶绿体类囊体膜 Chloroplast thylakoid membrane
ZmRUM1	生长素响应蛋白 Auxin- responsive protein	3	363	121	生长素激活信号通路 Auxin-activated signaling pathway DNA 模板转录的调控 Regulation of DNA- templated transcription	未知 Unknown	细胞核 Nucleolus
ZmTSAH1	色氨酸合成酶 Tryptophan synthase	1	1 020	340	色氨酸生物合成过程 Tryptophan biosynthetic process	色氨酸合成酶活性 Tryptophan synthase activity 吲哚-3-甘油-磷酸裂解酶活性 Indole-3-glycerol- phosphate lyase activity	叶绿体 Chloroplast 胞液 Cytosol
ZmNH4	nudix 水解酶 Nudix hydrolase 4	9	435	145	未知 Unknown	无机焦磷酸酶活性 Pyrophosphatase activity 水解酶活性 Hydrolase activity 金属离子结合 Metal ion binding	细胞质 Cytoplasm 细胞核 Nucleolus
ZmCL187-21A	60S核糖体蛋白L13 60S ribosomal protein L13	3	627	209	翻译 Translation	RNA结合 RNA binding 核糖体的结构成分 Structural constituent of ribosome	细胞质大核糖体亚基 Cytosolic large ribosomal subunit
ZmCCP2	半胱氨酸蛋白酶 Cysteine proteinase 2	7	1 083	361	免疫反应 Immune response 凋亡信号通路的正向调节 Positive regulation of apoptotic signaling pathway 参与蛋白质分解代谢过程的蛋白质水解 Proteolysis involved in protein catabolic process	半胱氨酸型内肽酶活性 Cysteine-type endopeptidase activity 苯丙氨酸转氨酶活性 Lyase activity	细胞外间隙 Extracellular space 溶酶体 Lysosome
ZmCSU27	紫外线-B-抑制蛋白 Ultraviolet-B- repressible protein	7	369	123	光合作用 Photosynthesis	蛋白质结构域特异性结合 Protein domain specific binding	细胞膜 Cytomembrane 叶绿体类囊体膜 Chloroplast thylakoid membrane
ZmNDF4	电子载体/电子传递体/铁离子结合蛋白 Electron carrier/ electron transporter/ iron ion binding protein	7	576	192	电子传递链 Electron transport chain 含P450的电子传递链 P450-containing electron transport chain 光系统Ⅰ中的光合电子传递 Photosynthetic electron transport in photosystem Ⅰ	2个铁原子, 2个硫原子簇结合 2 Iron, 2 sulfur cluster binding 电子传递活性 Electron transfer activity 金属离子结合 Metal ion binding 铁硫团簇结合 Iron-sulfur cluster binding	叶绿体类囊体膜 Chloroplast thylakoid membrane 胞液 Cytosol

玉米生物钟基因ZmPRR1-2互作蛋白质的筛选

Screening of proteins interacting with circadian clock gene ZmPRR1-2 in maize

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