Atypical OmpR/PhoB Subfamily Response Regulator GlnR of Actinomycetes Functions as a Homodimer, Stabilized by the Unphosphorylated Conserved Asp-focused Charge Interactions

doi:10.1074/jbc.M113.543504

CORC > 上海药物研究所 > 中国科学院上海药物研究所 > 药物发现与设计中心

	Atypical OmpR/PhoB Subfamily Response Regulator GlnR of Actinomycetes Functions as a Homodimer, Stabilized by the Unphosphorylated Conserved Asp-focused Charge Interactions
	Lin, Wei 5; Wang, Ying 4,5; Han, Xiaobiao 5; Zhang, Zilong 5; Wang, Chengyuan 2; Wang, Jin 5; Yang, Huaiyu 7; Lu, Yinhua 5; Jiang, Weihong 5; Zhao, Guo-Ping 1,3,4,5,6
刊名	JOURNAL OF BIOLOGICAL CHEMISTRY
	2014-05-30
卷号	289 期号:22 页码:15413-15425
关键词	Actinobacteria Crystal Structure Gene Regulation Phosphorylation Protein Conformation OmpR PhoB Subfamily Homodimer Receiver Domain Unphosphorylated Aspartate
ISSN号	0021-9258
DOI	10.1074/jbc.M113.543504
文献子类	Article
英文摘要	Background: Orphan response transcription factor GlnR regulates nitrogen metabolism in important actinomycetes. Results: GlnR has no typical phosphorylation pocket, where the only conserved Asp is unphosphorylated but is essential for functional homodimerization. Conclusion: Actinomycete GlnR is an atypical response regulator functioning as a homodimer. Significance: Conserved Asp-focused charge interactions of actinomycete GlnR are probably the mechanism that stabilizes the homodimer for physiological function. The OmpR/PhoB subfamily protein GlnR of actinomycetes is an orphan response regulator that globally coordinates the expression of genes related to nitrogen metabolism. Biochemical and genetic analyses reveal that the functional GlnR from Amycolatopsis mediterranei is unphosphorylated at the potential phosphorylation Asp(50) residue in the N-terminal receiver domain. The crystal structure of this receiver domain demonstrates that it forms a homodimer through the 4-5-5 dimer interface highly similar to the phosphorylated typical response regulator, whereas the so-called phosphorylation pocket is not conserved, with its space being occupied by an Arg(52) from the 3-3 loop. Both in vitro and in vivo experiments confirm that GlnR forms a functional homodimer via its receiver domain and suggest that the charge interactions of Asp(50) with the highly conserved Arg(52) and Thr(9) in the receiver domain may be crucial in maintaining the proper conformation for homodimerization, as also supported by molecular dynamics simulations of the wild type GlnR versus the deficient mutant GlnR(D50A). This model is backed by the distinct phenotypes of the total deficient GlnR(R52A/T9A) double mutant versus the single mutants of GlnR (i.e. D50N, D50E, R52A and T9A), which have only minor effects upon both dimerization and physiological function of GlnR in vivo, albeit their DNA binding ability is weakened compared with that of the wild type. By integrating the supportive data of GlnRs from the model Streptomyces coelicolor and the pathogenic Mycobacterium tuberculosis, we conclude that the actinomycete GlnR is atypical with respect to its unphosphorylated conserved Asp residue being involved in the critical Arg/Asp/Thr charge interactions, which is essential for maintaining the biologically active homodimer conformation.
资助项目	National Basic Research Program of China[2012CB721102] ; National Nature Science Foundation[31322016] ; National Nature Science Foundation[31121001] ; National Nature Science Foundation[30830002] ; Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences[2012OHTP] ; Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences[2009CSP001] ; Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences[KSCX2-EW-J-12]
WOS关键词	2-COMPONENT SIGNAL-TRANSDUCTION ; AMYCOLATOPSIS-MEDITERRANEI U32 ; STREPTOMYCES-COELICOLOR A3(2) ; MYCOBACTERIUM-TUBERCULOSIS ; NITROGEN-METABOLISM ; RECEIVER DOMAIN ; ACTIVATION ; MECHANISM ; PHOSPHORYLATION ; TRANSCRIPTION
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
WOS记录号	WOS:000337465400025
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/277065]
专题	药物发现与设计中心
通讯作者	Zhao, Guo-Ping
作者单位	1.Chinese Univ Hong Kong, Prince Wales Hosp, Li Ka Shing Inst Hlth Sci, Shatin, Hong Kong, Peoples R China; 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, State Key Lab Plant Mol Genet, Shanghai 200032, Peoples R China; 3.Chinese Natl Human Genome Ctr Shanghai, Shanghai MOST Key Lab Dis & Hlth Genom, Shanghai 201203, Peoples R China; 4.Fudan Univ, Sch Life Sci, Dept Microbiol & Microbial Engn, State Key Lab Genet Engn, Shanghai 200433, Peoples R China; 5.Chinese Acad Sci, Key Lab Synthet Biol, Shanghai 200032, Peoples R China; 6.Chinese Univ Hong Kong, Prince Wales Hosp, Dept Microbiol, Shatin, Hong Kong, Peoples R China; 7.Chinese Acad Sci, Inst Mat Med, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Lin, Wei,Wang, Ying,Han, Xiaobiao,et al. Atypical OmpR/PhoB Subfamily Response Regulator GlnR of Actinomycetes Functions as a Homodimer, Stabilized by the Unphosphorylated Conserved Asp-focused Charge Interactions[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2014,289(22):15413-15425.
APA	Lin, Wei.,Wang, Ying.,Han, Xiaobiao.,Zhang, Zilong.,Wang, Chengyuan.,...&Zhang, Peng.(2014).Atypical OmpR/PhoB Subfamily Response Regulator GlnR of Actinomycetes Functions as a Homodimer, Stabilized by the Unphosphorylated Conserved Asp-focused Charge Interactions.JOURNAL OF BIOLOGICAL CHEMISTRY,289(22),15413-15425.
MLA	Lin, Wei,et al."Atypical OmpR/PhoB Subfamily Response Regulator GlnR of Actinomycetes Functions as a Homodimer, Stabilized by the Unphosphorylated Conserved Asp-focused Charge Interactions".JOURNAL OF BIOLOGICAL CHEMISTRY 289.22(2014):15413-15425.