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小肠结肠炎耶尔森菌FlhD和FlhC的基因芯片分析:影响氨甲酰磷酸合成与降解的酶的调控

Gene array analysis of Yersinia enterocolitica FlhD and FlhC: regulation of enzymes affecting synthesis and degradation of carbamoylphosphate.

作者信息

Kapatral Vinayak, Campbell John W, Minnich Scott A, Thomson Nicholas R, Matsumura Philip, Prüß Birgit M

机构信息

Integrated Genomics, Inc., 2201 West Campbell Park Dr., Chicago, IL 60612, USA.

Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83843, USA.

出版信息

Microbiology (Reading). 2004 Jul;150(Pt 7):2289-2300. doi: 10.1099/mic.0.26814-0.

Abstract

This paper focuses on global gene regulation by FlhD/FlhC in enteric bacteria. Even though Yersinia enterocolitica FlhD/FlhC can complement an Escherichia coli flhDC mutant for motility, it is not known if the Y. enterocolitica FlhD/FlhC complex has an effect on metabolism similar to E. coli. To study metabolic gene regulation, a partial Yersinia enterocolitica 8081c microarray was constructed and the expression patterns of wild-type cells were compared to an flhDC mutant strain at 25 and 37 degrees C. The overlap between the E. coli and Y. enterocolitica FlhD/FlhC regulated genes was 25 %. Genes that were regulated at least fivefold by FlhD/FlhC in Y. enterocolitica are genes encoding urocanate hydratase (hutU), imidazolone propionase (hutI), carbamoylphosphate synthetase (carAB) and aspartate carbamoyltransferase (pyrBI). These enzymes are part of a pathway that is involved in the degradation of L-histidine to L-glutamate and eventually leads into purine/pyrimidine biosynthesis via carbamoylphosphate and carbamoylaspartate. A number of other genes were regulated at a lower rate. In two additional experiments, the expression of wild-type cells grown at 4 or 25 degrees C was compared to the same strain grown at 37 degrees C. The expression of the flagella master operon flhD was not affected by temperature, whereas the flagella-specific sigma factor fliA was highly expressed at 25 degrees C and reduced at 4 and 37 degrees C. Several other flagella genes, all of which are under the control of FliA, exhibited a similar temperature profile. These data are consistent with the hypothesis that temperature regulation of flagella genes might be mediated by the flagella-specific sigma factor FliA and not the flagella master regulator FlhD/FlhC.

摘要

本文聚焦于肠道细菌中FlhD/FlhC对全局基因的调控。尽管小肠结肠炎耶尔森菌的FlhD/FlhC能够互补大肠杆菌flhDC突变体的运动能力,但尚不清楚小肠结肠炎耶尔森菌的FlhD/FlhC复合物是否具有与大肠杆菌类似的对代谢的影响。为了研究代谢基因调控,构建了部分小肠结肠炎耶尔森菌8081c微阵列,并比较了野生型细胞与flhDC突变株在25℃和37℃下的表达模式。大肠杆菌和小肠结肠炎耶尔森菌中受FlhD/FlhC调控的基因重叠率为25%。在小肠结肠炎耶尔森菌中受FlhD/FlhC至少五倍调控的基因包括编码尿刊酸水合酶(hutU)、咪唑啉丙酸酶(hutI)、氨甲酰磷酸合成酶(carAB)和天冬氨酸氨甲酰转移酶(pyrBI)的基因。这些酶是参与将L-组氨酸降解为L-谷氨酸的途径的一部分,并最终通过氨甲酰磷酸和氨甲酰天冬氨酸进入嘌呤/嘧啶生物合成。其他一些基因的调控率较低。在另外两个实验中,比较了在4℃或25℃下生长的野生型细胞与在37℃下生长的同一菌株的表达情况。鞭毛主操纵子flhD的表达不受温度影响,而鞭毛特异性σ因子fliA在25℃时高表达,在4℃和37℃时降低。其他几个均受FliA控制的鞭毛基因也表现出类似的温度分布。这些数据与以下假设一致,即鞭毛基因的温度调控可能由鞭毛特异性σ因子FliA介导,而非鞭毛主调节因子FlhD/FlhC。

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