IDENTIFICATION OF SEQUENCES RESPONSIBLE FOR TRANSCRIPTIONAL REGULATION OF THE STRONGLY EXPRESSED METHANOL OXIDASE-ENCODING GENE IN HANSENULA-POLYMORPHA
S. Godecke et al., IDENTIFICATION OF SEQUENCES RESPONSIBLE FOR TRANSCRIPTIONAL REGULATION OF THE STRONGLY EXPRESSED METHANOL OXIDASE-ENCODING GENE IN HANSENULA-POLYMORPHA, Gene, 139(1), 1994, pp. 35-42
The methylotrophic yeasts have been the subject of intensive studies,
because of their highly regulated methanol metabolism and the biogenes
is of peroxisomes. We investigated the 5' regulatory region of the MOX
gene from the yeast, Hansemula polymorpha, encoding the peroxisomal m
ethanol oxidase, the key enzyme of methanol metabolism. This tightly r
egulated yeast promoter of approximately 1.5 kb is unusually large, an
d also of remarkable strength under inducing conditions, belonging to
the strongest yeast promoters yet described. Deletion analyses reveale
d a complex promoter structure composed of several sequence elements w
ith positive and negative regulatory effects on reporter gene expressi
on and a pronounced cooperation between the elements. Specific binding
of several factors was detected in vitro by gel retardation and DNase
I footprinting experiments. On the basis of deletion data, two bindin
g sites could be identified as upstream activation sequences (UAS1 and
UAS2) and one binding site as an upstream repressing sequence (URS1).