A chimeric light-regulated amino acid transport system allows the isolation of blue light regulator (blr) mutants of Neurospora crassa.
We have developed a system for the isolation of Neurospora crassa mutants that shows altered responses to blue light. To this end we have used the light-regulated promoter of the albino-3 gene fused to the neutral amino acid permease gene mtr. The product of the mtr gene is required for the uptake of neutral aliphatic and aromatic amino acids, as well as toxic analogs such as p-flurophenylalanine or 4-methyltryptophan. mtr trp-2-carrying cells were transformed with the al-3 promoter-mtr wild-type gene (al-3p-mtr+) to obtain a strain with a light-regulated tryptophan uptake. This strain is sensitive to p-fluorophenylalanine when grown under illumination and resistant when grown in the dark. UV mutagenesis of the al-3p-mtr(+)-carrying strain allowed us to isolate two mutant strains, BLR-1 and BLR-2 (blue light regulator), that are light-resistant to p-fluorophenylalanine and have lost the ability to grow on tryptophan. These two strains have a pale-orange phenotype and show down-regulation of all the photoregulated genes tested (al-3, al-1, con-8, and con-10). Mutations in the BLR strains are not allelic with white collar 1 or white collar 2, regulatory genes that are also involved in the response to blue light.
Carattoli
A
A
Dipartimento di Biopatologia Umana, Università di Roma La Sapienza, Rome, Italy.
Kato
E
E
Rodriguez-Franco
M
M
Stuart
W D
WD
Macino
G
G
D026905:Amino Acid Transport Systems; D000596:Amino Acids / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D002678:Chimera; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis; D015966:Gene Expression Regulation, Fungal / Q000528:radiation effects; D005800:Genes, Fungal; D008027:Light; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009492:Neurospora crassa / Q000235:genetics* / Q000378:metabolism* / Q000528:radiation effects; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D014466:Ultraviolet Rays
D026905:Amino Acid Transport Systems; D000596:Amino Acids; D002352:Carrier Proteins; D017931:DNA Primers; D004271:DNA, Fungal; D012333:RNA, Messenger