Inability of Ogataea parapolymorpha pho91-Δ mutant to produce active methanol oxidase can be compensated by inactivation of the PHO87 gene.

Abstract

Cells of the methylotrophic yeast Ogataea parapolymorpha have two genes encoding low-affinity phosphate transporters: PHO87, encoding the plasma membrane transporter, and PHO91, encoding a protein, which is homologous to the Saccharomyces cerevisiae vacuolar membrane transporter. Earlier, we reported that inactivation of PHO91 in O. parapolymorpha interferes with methanol utilization due to the lack of activity of methanol oxidase encoded by the MOX gene. In this work, we showed that this defect was completely suppressed by inactivating the PHO87 gene or introducing additional copies of the MOX gene into the cell. The PHO91 gene knockout decreased the level of long-chained polyphosphates only in methanol-grown cells, but not in glucose-grown cells. This effect remained even in the strain with extra copies of MOX, which rescues the ability of the mutant to grow on methanol. In contrast, the PHO87 gene knockout changed the levels of short-chained and long-chained polyphosphates in both methanol- and glucose-grown cells. Inactivation of PHO91 did not change vanadate resistance, while inactivation of PHO87 increased this resistance. Our data suggest that in O. parapolymorpha, Pho87 and Pho91 transporters have different roles in inorganic polyphosphate metabolism and adaptation to methanol consumption.