Variation in transport explains polymorphism of histidine and urocanate utilization in a natural Pseudomonas population.

Variation in transport explains polymorphism of histidine and urocanate utilization in a natural Pseudomonas population.
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运输的变化解释了自然假单胞菌群体中组氨酸和尿刊酸利用的多态性。

DOI:
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发表时间:
2012
影响因子:
5.1
通讯作者:
P. Rainey
P. Rainey
中科院分区:
生物学2区
文献类型:
--
作者:
Xue‐Xian Zhang;Hao Chang;Sieu L. Tran;J. Gauntlett;G. Cook;P. Rainey

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表型变异是自然选择进化的基本要求。虽然在自然种群中表型变异的证据比比皆是,但其遗传基础却很少被理解。在这里,我们报告的变化,在植物定植假单胞菌利用组氨酸,及其衍生物,尿刊酸,作为唯一的碳和氮源的能力。从164个叶圈定殖假单胞菌菌株的群体中,77%能够利用组氨酸和尿刊酸(His(+),Uro(+))作为生长底物,而其余的可以利用组氨酸,但不能利用尿刊酸(His(+),Uro(-)),反之亦然(His(-),Uro(+))。一个在电脑分析的小屋位点,确定能力,利用组氨酸和尿刊酸,从基因组测序的假单胞菌菌株,显示出显着的变化,推定的转运蛋白的数量。为了确定转运蛋白基因特异性组氨酸和尿刊酸,我们专注于一个单一的基因型荧光假单胞菌,菌株SBW 25,这是能够利用这两种底物。定点诱变结合[(3)H]组氨酸转运试验表明,hutT(u)编码尿刊酸特异性转运蛋白; hutT(h)编码主要的高亲和力组氨酸转运蛋白; hutXWV编码ABC型转运蛋白,在组氨酸摄取中起次要作用。将来自SBW 25的hutT(h)和hutT(u)的克隆拷贝引入不能利用组氨酸或尿刊酸的菌株中,补充了缺陷,证明这些菌株中缺乏功能性转运蛋白。总之,我们的数据表明,运输系统的变化,而不是在代谢基因,解释了自然发生的表型多态性。
Phenotypic variation is a fundamental requirement for evolution by natural selection. While evidence of phenotypic variation in natural populations abounds, its genetic basis is rarely understood. Here we report variation in the ability of plant-colonizing Pseudomonas to utilize histidine, and its derivative, urocanate, as sole sources of carbon and nitrogen. From a population of 164 phyllosphere-colonizing Pseudomonas strains, 77% were able to utilize both histidine and urocanate (His(+) , Uro(+) ) as growth substrates, whereas the remainder could utilize histidine, but not urocanate (His(+) , Uro(-) ), or vice versa (His(-) , Uro(+) ). An in silico analysis of the hut locus, which determines capacity to utilize both histidine and urocanate, from genome-sequenced Pseudomonas strains, showed significant variation in the number of putative transporters. To identify transporter genes specific for histidine and urocanate, we focused on a single genotype of Pseudomonas fluorescens, strain SBW25, which is capable of utilizing both substrates. Site-directed mutagenesis, combined with [(3) H]histidine transport assays, shows that hutT(u) encodes a urocanate-specific transporter; hutT(h) encodes the major high-affinity histidine transporter; and hutXWV encodes an ABC-type transporter that plays a minor role in histidine uptake. Introduction of cloned copies of hutT(h) and hutT(u) from SBW25 into strains incapable of utilizing either histidine, or urocanate, complemented the defect, demonstrating a lack of functional transporters in these strains. Taken together our data show that variation in transport systems, and not in metabolic genes, explains a naturally occurring phenotypic polymorphism.
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