IMMUNOLOGICAL DETECTION AND SUBCELLULAR LOCALIZATION OF THE GENERAL PURINE TRANSPORTER UAPC FROM ASPERGILLUS NIDULANS

VALDEZ TAUBAS J; DIALLINAS, G; SCAZZOCCHIO C; ROSA AL

FUNGAL GENETICS AND BIOLOGY

ACADEMIC PRESS INC ELSEVIER SCIENCE

Año: 2000 vol. 30 p. 105 - 113

1087-1845

The uapC gene of Aspergillus nidulans belongs to a family of nucleobase-specific transporters conserved in prokaryotic and eucaryotic organisms. We report the use of immunological and green fluorescent protein based strategies to study protein expression and subcellular distribution of UapC. A chimeric protein containing a plant-adapted green fluorescent protein (sGFP) fused to the C-terminus of UapC was shown to be functional in vivo, as it complements a triple mutant (i.e., uapC− uapA− azgA−) unable to grow on uric acid as the sole nitrogen source. UapC-GFP is located in the plasma membrane and, secondarily, in internal structures observed as fluorescent dots. A strong correlation was found between cellular levels of UapC-GFP fluorescence and known patterns of uapC gene expression. This work represents the first in vivo study of protein expression and subcellular localization of a filamentous fungal nucleobase transporter.