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酵母碳酸氢盐转运蛋白同源物Bor1p的转运及调控特性

Transport and regulatory characteristics of the yeast bicarbonate transporter homolog Bor1p.

作者信息

Jennings Michael L, Howren Todd R, Cui Jian, Winters Maria, Hannigan Robyn

机构信息

Dept. of Physiology and Biophysics, University of Arkansas for Medical Sciences, 4301 W. Markham St., Mail Slot 505, Little Rock, AR 72205, USA.

出版信息

Am J Physiol Cell Physiol. 2007 Jul;293(1):C468-76. doi: 10.1152/ajpcell.00286.2005. Epub 2007 Apr 25.

Abstract

The functional properties of the Saccharomyces cerevisiae bicarbonate transporter homolog Bor1p (YNL275wp) were characterized by measuring boron (H(3)BO(3)), Na(+), and Cl(-) fluxes. Neither Na(+) nor Cl(-) appears to be a transported substrate for Bor1p. Uphill efflux of boron mediated by Bor1p was demonstrated directly by loading cells with boron and resuspending in a low-boron medium. Cells with intact BOR1, but not the deletant strain, transport boron outward until the intracellular concentration is sevenfold lower than that in the medium. Boron efflux through Bor1p is a saturable function of intracellular boron (apparent K(m) approximately 1-2 mM). The extracellular pH dependences of boron distribution and efflux indicate that uphill efflux is driven by the inward H(+) gradient. Addition of 30 mM HCO(3)(-) does not affect boron extrusion by Bor1p, indicating that HCO(3)(-) does not participate in Bor1p function. Functional Bor1p is present in cells grown in medium with no added boron, and overnight growth in 10 mM H(3)BO(3) causes only a small increase in the levels of functional Bor1p and in BOR1 mRNA. The fact that Bor1p is expressed when there is no need for boron extrusion and is not strongly induced in the presence of growth-inhibitory boron concentrations is surprising if the main physiological function of yeast Bor1p is boron efflux. A possible role in vacuolar dynamics for Bor1p was recently reported by Decker and Wickner. Under the conditions used presently, there appears to be mildly abnormal vacuolar morphology in the deletant strain.

摘要

通过测量硼(H₃BO₃)、Na⁺和Cl⁻通量,对酿酒酵母碳酸氢盐转运蛋白同源物Bor1p(YNL275wp)的功能特性进行了表征。Na⁺和Cl⁻似乎都不是Bor1p的转运底物。通过用硼加载细胞并重悬于低硼培养基中,直接证明了由Bor1p介导的硼的上坡外流。具有完整BOR1的细胞,而不是缺失菌株,将硼向外转运,直到细胞内浓度比培养基中的浓度低七倍。通过Bor1p的硼外流是细胞内硼的饱和函数(表观Kₘ约为1 - 2 mM)。硼分布和外流的细胞外pH依赖性表明,上坡外流是由内向H⁺梯度驱动的。添加30 mM HCO₃⁻不会影响Bor1p对硼的挤出,表明HCO₃⁻不参与Bor1p的功能。功能性Bor1p存在于未添加硼的培养基中生长的细胞中,在10 mM H₃BO₃中过夜生长仅导致功能性Bor1p水平和BOR₁ mRNA水平略有增加。如果酵母Bor1p的主要生理功能是硼外流,那么在不需要硼挤出时Bor1p就表达,并且在存在生长抑制性硼浓度时不会被强烈诱导,这一事实令人惊讶。Decker和Wickner最近报道了Bor1p在液泡动力学中的可能作用。在目前使用的条件下,缺失菌株中似乎存在轻度异常的液泡形态。

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