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J. Biol. Chem., Vol. 279, Issue 30, 31937-31942, July 23, 2004

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Glycerophosphoinositol, a Novel Phosphate Source Whose Transport Is Regulated by Multiple Factors in Saccharomyces cerevisiae^*

Claudia Almaguer, Wei Cheng, Christi Nolder, and Jana Patton-Vogt

From the Department of Biological Sciences, Duquesne University, Pittsburgh, Pennsylvania 15282

Git1p mediates the transport of the phospholipid metabolite, glycerophosphoinositol, into Saccharomyces cerevisiae. We report that phosphate limitation and inositol limitation affect GIT1 expression and Git1p transport activity via distinct mechanisms that involve multiple transcription factors. GIT1 transcript levels and Git1p activity are greater in cells starved for phosphate, with or without inositol limitation, than in cells only limited for inositol. Furthermore, the kinetics of GIT1 transcript accumulation and Git1p activity upon transfer of cells to phosphate starvation media are different from those obtained upon transfer of cells to inositol-free media. Pho2p and Pho4p are required for GIT1 expression and for Git1p transport activity under all growth conditions tested. In contrast, Ino2p and Ino4p are required for full GIT1 expression when inositol is limiting, with or without phosphate limitation, but not when only phosphate is limiting. Greatly reduced transport activity was detected in ino2 and ino4 cells under all growth conditions. A 300-base pair region of the GIT1 promoter containing potential Pho4p binding sites was shown to be required for full GIT1 expression. Git1p appears to act as a H⁺-symporter, and neither inositol nor phosphate effectively compete with glycerophosphoinositol for transport by Git1p. Glycerophosphoinositol was shown previously to support the growth of an inositol auxotroph. Remarkably, we now report that glycerophosphoinositol can act as the sole source of phosphate for the cell, providing functional relevance for the regulation of Git1p transport activity by phosphate.

Received for publication, April 1, 2004 , and in revised form, May 11, 2004.

^* This work was supported by Grant GM59817 from the National Institutes of Health (to J. P.-V.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 412-396-1053; Fax: 412-396-5907; E-mail: pattonvogt{at}duq.edu.

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