An iNTT system for the large-scale screening of differentially expressed, nuclear-targeted proteins: cold-treatment-induced nucleoproteins in Rye (Secale cereale L.)

doi:10.1186/s12864-016-2548-y

	An iNTT system for the large-scale screening of differentially expressed, nuclear-targeted proteins: cold-treatment-induced nucleoproteins in Rye (Secale cereale L.)
	Cao, Xinyou 1,2,3,4,5; Chen, Xueyan 3,4,5,6; Liu, Yangna 1,2; Xu, Zhaoshi 1,2; Li, Liancheng 1,2; Zhou, Yongbin 1,2; Liu, Jianjun 3,4,5; Zhao, Zhendong 3,4,5; Chen, Ming 1,2; Ma, Youzhi 1,2
刊名	BMC GENOMICS
	2016
卷号	17 页码:-
关键词	Nuclear-targeted protein iNTT system After suppression subtraction method Low-temperature stress Rye
ISSN号	1471-2164
DOI	10.1186/s12864-016-2548-y
通讯作者	Cao, Xinyou
英文摘要	Background: Nuclear proteins play critical roles in regulating mRNA transcription and processing, DNA replication, and epigenetic genome modification. Therefore, the ability to monitor changes in nuclear proteins is helpful not only to identify important regulatory proteins but also to study the mechanisms of actions of nuclear proteins. However, no effective methods have been developed yet. Rye is strongly resistant to various biotic and abiotic stresses; however, few genes have been functionally characterized to date due to the complexity of its genome and a lack of genomic sequence information.Results: We developed an integrative Nuclear Transportation Trap (iNTT) system that includes an improved nuclear transportation trap and utilizes the "after suppression subtraction" method. Oligonucleotides encoding a nuclear localization signal (NLS) or a transcription factor, GmAREB, were inserted into pLexAD or pLexAD-NES, respectively, and then transformed into yeast cells (EGY48). We showed that the pLexAD vector expressing a cDNA library in the iNTT system was more efficient for screening than the vector pLexAD-NES, which has previously been used in an NTT system. We used the iNTT system to screen a cDNA library of cold-treated rye. A total of 241 unique genes were identified, including 169 differentially expressed proteins; of these, 106 were of known and 63 were of unknown function. Moreover, 82 genes (49 %) among the 169 differentially expressed genes were predicted to contain an NLS domain. Thirty-three (31 %) of the 106 functionally known proteins have DNA-binding activity. To test the specificity of the nuclear proteins identified using the iNTT screen, four of the proteins differentially expressed in response to temperature stress, ScT1 (a heat shock protein), ScT36 (a MYB-like transcription factor), ScT133 (an ERF-like transcription factor) and ScT196 (a protein of unknown function), were studied in more depth. These proteins were shown to exclusively localize to the nucleus, and their expression levels were increased in response to low-temperature stress. To identify the function of these screened nuclear proteins, ScT1-and ScT36-transgenic Arabidopsis plants were constructed, and ScT1 or ScT36 overexpression was found to enhance tolerance to high-temperature or freezing stresses, respectively.Conclusions: The newly developed iNTT system provides an effective method for identifying nuclear-targeted proteins and monitoring induced expression levels. ScT1 and ScT36 might be good candidate genes for improving the stress tolerance of plants by genetic transformation.
学科主题	Biotechnology & Applied Microbiology ; Genetics & Heredity
语种	英语
出版者	BIOMED CENTRAL LTD
WOS记录号	WOS:000371595300004
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/4563]
专题	作物科学研究所_分子生物学系
作者单位	1.Minist Agr, Key Lab Crop Genet & Breeding, Natl Key Facil Crop Genet Resources & Genet Impro, Beijing 100081, Peoples R China 2.Chinese Acad Agr Sci, Inst Crop Sci, Beijing 100081, Peoples R China 3.Shandong Acad Agr Sci, Crop Res Inst, Jinan 250100, Peoples R China 4.Natl Engn Lab Wheat & Maize, Jinan 250100, Peoples R China 5.Minist Agr, Key Lab Wheat Biol & Genet Improvement North Yell, Jinan 250100, Peoples R China 6.Northwest A&F Univ, Coll Agron, Yangling 712100, Peoples R China
推荐引用方式 GB/T 7714	Cao, Xinyou,Chen, Xueyan,Liu, Yangna,et al. An iNTT system for the large-scale screening of differentially expressed, nuclear-targeted proteins: cold-treatment-induced nucleoproteins in Rye (Secale cereale L.)[J]. BMC GENOMICS,2016,17:-.
APA	Cao, Xinyou.,Chen, Xueyan.,Liu, Yangna.,Xu, Zhaoshi.,Li, Liancheng.,...&Ma, Youzhi.(2016).An iNTT system for the large-scale screening of differentially expressed, nuclear-targeted proteins: cold-treatment-induced nucleoproteins in Rye (Secale cereale L.).BMC GENOMICS,17,-.
MLA	Cao, Xinyou,et al."An iNTT system for the large-scale screening of differentially expressed, nuclear-targeted proteins: cold-treatment-induced nucleoproteins in Rye (Secale cereale L.)".BMC GENOMICS 17(2016):-.