细菌GntR家族转录调控因子的研究进展

GntR家族转录调控因子是细菌中分布最为广泛的一类螺旋–转角–螺旋(helix-turn-helix,HTH)转录调控因子,此家族转录调控因子包含两个功能域,分别是N端的DNA结合结构域和C端的效应物结合结构域/寡聚化作用结构域。DNA结合结构域的氨基酸序列是非常保守的,但效应物结合结构域/寡聚化作用结构域的氨基酸序列却存在很大的差异性。目前许多GntR家族的转录调控因子已经被鉴定,这些转录因子调控细菌许多不同的细胞过程,如运动性、葡萄糖代谢、细菌的耐药性、病原细菌的致病力等。本文主要阐述了GntR家族转录调控因子的发现、二级结构、生物学功能、调控模式等方面的研究进展,旨在为研究者全面、深入地了解GntR家族转录调控因子的功能及作用机理提供帮助。

十字花科黑腐病菌中转录因子HpaR1与Clp调控一个糖苷水解酶基因表达的分析

十字花科黑腐病菌(Xanthomonas campestris pv. campestris, Xcc)是一种维管束致病菌,能够引起寄主的黑腐病,是研究植物病原细菌与植物互作的一种重要模式菌株。在Xcc中,GntR家族的全局性转录调控因子HpaR1参与调控Xcc的运动、胞外多糖和胞外酶的合成等许多细胞过程,并与Xcc的过敏反应(hypersensitive response,HR)和致病相关;全局性转录调控因子Clp则参与调控胞外酶和胞外多糖的分泌与合成,并与黄单胞菌的致病相关。前期研究发现,Xcc中的转录调控因子HpaR1和Clp均能与糖苷水解酶(glycoside hydrolase)编码基因(命名为ghy基因)的启动子区结合。为探究转录调控因子HpaR1和Clp共同调控ghy基因表达的分子机理,本研究首先通过凝胶阻滞分析(electrophoresis mobility shift assay,EMSA)发现HpaR1和Clp在体外能够结合在ghy基因的启动子区;利用染色质免疫共沉淀(chromatin immunoprecipitation, ChIP)方法,进一步证实HpaR1和Clp在细胞内能够结合在ghy基因启动子区。通过5’-cDNA末端快速扩增(rapid amplication of 5’-cDNAends,5’-RACE)和DNase I保护实验(DNase I footprinting)确定HpaR1和Clp均结合在ghy基因启动子的–35区上游,并且HpaR1的结合位点位于Clp结合位点的上游。通过实时荧光定量PCR(real time fluorescence quantitative PCR,RT-qPCR)和体外转录的方法,发现HpaR1抑制ghy基因的转录,而Clp激活ghy基因的转录。当二者共同存在时,HpaR1能够抑制Clp对ghy基因的转录激活作用。HpaR1和Clp单独存在时,分别负调控和正调控ghy基因的转录,推测HpaR1尽管位于ghy基因启动子-35区上游,但可能通过抑制RNA聚合酶的活性来调控ghy基因的表达。

Variation in plant diversity and dominance across dune fixation stages in the Chinese steppe zone

Aims Our aim was to study how diversity and dominance of plant species and plant functional types(PFTs)change and covary across three dune fixation stages in the Chinese steppe zone.Methods In the Chinese steppe zone,we measured coverage,mean height and density of each plant species in three types of dunes(mobile,semi-fixed and fixed dunes)in four sites(Mu Us,Otindag,Hulunbeir and Horqin).Plant species were grouped into 24 PFTs according to their lifespan,photosynthetic pathway,reproductive mode and life form.Dominance of each plant species and PFT were determined,and species diversity and PFT diversity were quantified using Shannon–Wiener index.Important Findings PFT diversity was positively related to plant species diversity in each dune stage,but PFT diversity increased more with increasing plant species diversity in the mobile and semi-fixed dunes than in the fixed dunes.Dune fixation stage explained 87.2%of the variation in plant species diversity and 84.8%of the variation in PFT diversity.Dominant species and PFTs differed among the three dune fixation stages;the more fixed the dunes were,the more perennial,shrubby,clonal and C3 species co-dominated.Specifically,in mobile dunes annual C4 non-clonal herbs were the most dominant,and in semi-fixed and fixed dunes perennial C3 clonal shrubs were most dominant.