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Abstract

Small peptides (2–5 amino acid residues) are transported into via two transport systems: PTR (eptide ansport) for di-/tripeptides and OPT (ligoeptide ransport) for oligopeptides of 4–5 amino acids in length. Although regulation of the PTR system has been studied in some detail, neither the regulation of the OPT family nor the environmental conditions under which family members are normally expressed have been well studied in . Using a reporter gene construct fused to 1 kb DNA from upstream of the genes and , which encode the two oligopeptide transporters, the relative expression levels of these genes were measured in a variety of environmental conditions. Uptake assays were also conducted to measure functional protein levels at the plasma membrane. It was found that was up-regulated in sulfur-free medium, and that Ptr3p and Ssy1p, proteins involved in regulating the di-/tripeptide transporter encoding gene via amino acid sensing, were required for expression in a sulfur-free environment. In contrast, as measured by response to toxic tetrapeptide and by real-time PCR, was not regulated through Cup9p, which is a repressor for expression, although Cup9p did repress expression. In addition, all of the 20 naturally occurring amino acids, except the sulfur-containing amino acids methionine and cysteine, up-regulated , with the greatest change in expression observed when cells were grown in sulfur-free medium. These data demonstrate that regulation of the OPT system has both similarities and differences to regulation of the PTR system, allowing the yeast cell to adapt its utilization of small peptides to various environmental conditions.

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2006-10-01
2019-11-18
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vol. , part 10, pp. 3133-3145

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Media formulations. Synthetic complete medium (SC), with modifications made for media containing reduced carbon (r C) or no carbon (no C), reduced nitrogen (r N) or nitrogen (no N), reduced sulfur (r S) or no sulfur (no S), and reduced nitrogen and sulfur (r NS) or no nitrogen and sulfur (no NS). The SC formula was first referred to as ‘nitrogen base for carbon assimilation tests’ but here includes no amino acids, and it was assembled as described by Wickerham (1946, 1951). Compounds listed are in mg/l.

Amino acids added to satisfy auxotrophic requirements or to test induction of OPT1.



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