Isolation of mutants of Hansenula polymorpha defective in the assembly of octameric alcohol oxidase.
Alcohol oxidase (AO) is a peroxisomal enzyme that catalyses the first step in methanol metabolism in yeast. Monomeric, inactive AO protein is synthesised in the cytosol and subsequently imported into peroxisomes, where the enzymatically active, homo-octameric form is found. The mechanisms involved in AO octamer assembly are largely unclear. Here we describe the isolation of Hansenula polymorpha mutants specifically affected in AO assembly. These mutants are unable to grow on methanol and display reduced AO activities. Based on their phenotypes, three major classes of mutants were isolated. Three additional mutants were isolated that each displayed a unique phenotype. Complementation analysis revealed that the isolated AO assembly mutants belonged to 10 complementation groups.
van Dijk
Ralf
R
Eukaryotic Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, Biological Centre, University of Groningen, 9750 AA Haren, The Netherlands.
Lahchev
Kancho L
KL
Kram
Anita M
AM
van der Klei
Ida J
IJ
Veenhuis
Marten
M
D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005816:Genetic Complementation Test; D000432:Methanol / Q000378:metabolism; D008854:Microscopy, Electron; D009154:Mutation; D020675:Peroxisomes / Q000378:metabolism; D010843:Pichia / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000648:ultrastructure
D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D000432:Methanol