Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involv...Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involved in autoimmunity initiated by type one protein phosphatase 4 mutation (topp4-1) in Arabidopsis, however, its role in planta is still unclear. This study employed Nicotiana benthamiana, a model platform, to conduct an overall structural and functional analysis of SUT1 protein. The transient expression results revealed that SUT1 is a typical CNL (CC-NBS-LRR) receptor, both fluorescence data and biochemical results showed the protein is mainly anchored on the plasma membrane due to its N-terminal acylation site. Further truncation experiments announced that its CC (coiled-coil) domain possessed cell-death-inducing activity. The outcomes of point mutations analysis revealed that not only the CC domain, but also the full-length SUT1 protein, whose function and subcellular localization are influenced by highly conserved hydrophobic residues. These research outcomes provided favorable clues for elucidating the activation mechanism of SUT1.展开更多
根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此...根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此基因被命名为GHNBS。该基因的编码区长2 583 bp,编码861个氨基酸,GHNBS编码的氨基酸序列与拟南芥R基因具有28%的同源性。GHNBS与拟南芥的其他几个NBS-LRR基因比较发现,它们在保守区外的相似性相当低。Southern杂交和网上数据库搜索分析都表明GHNBS是1个寡拷贝基因。通过半定量RT-PCR分析发现GHNBS在棉花的蕾、花瓣、韧皮部、根及叶中均有表达且在根、叶中表达量比其它部位强,在木质部基本不表达。展开更多
文摘Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involved in autoimmunity initiated by type one protein phosphatase 4 mutation (topp4-1) in Arabidopsis, however, its role in planta is still unclear. This study employed Nicotiana benthamiana, a model platform, to conduct an overall structural and functional analysis of SUT1 protein. The transient expression results revealed that SUT1 is a typical CNL (CC-NBS-LRR) receptor, both fluorescence data and biochemical results showed the protein is mainly anchored on the plasma membrane due to its N-terminal acylation site. Further truncation experiments announced that its CC (coiled-coil) domain possessed cell-death-inducing activity. The outcomes of point mutations analysis revealed that not only the CC domain, but also the full-length SUT1 protein, whose function and subcellular localization are influenced by highly conserved hydrophobic residues. These research outcomes provided favorable clues for elucidating the activation mechanism of SUT1.
文摘根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此基因被命名为GHNBS。该基因的编码区长2 583 bp,编码861个氨基酸,GHNBS编码的氨基酸序列与拟南芥R基因具有28%的同源性。GHNBS与拟南芥的其他几个NBS-LRR基因比较发现,它们在保守区外的相似性相当低。Southern杂交和网上数据库搜索分析都表明GHNBS是1个寡拷贝基因。通过半定量RT-PCR分析发现GHNBS在棉花的蕾、花瓣、韧皮部、根及叶中均有表达且在根、叶中表达量比其它部位强,在木质部基本不表达。
