The Hansenula polymorpha per6 mutant is affected in two adjacent genes which encode dihydroxyacetone kinase and a novel protein, Pak1p, involved in peroxisome integrity.
The Hansenula polymorpha per6-210 mutant is impaired in respect of growth on methanol (Mut-) and is characterized by aberrant peroxisome formation. The functionally complementing DNA fragment contains two open reading frames. The first encodes dihydroxyacetone kinase (DAK), a cytosolic enzyme essential for formaldehyde assimilation; the second ORF codes for a novel protein (Pak1p). We have demonstrated that per6-210 cells lack DAK activity, causing the Mut- phenotype, and have strongly reduced levels of Pak1p, resulting in peroxisomal defects. Sequence analysis revealed that per6-210 contains a mutation in the 3' end of the DAK coding region, which overlaps with the promoter region of PAK1. Possibly this mutation also negatively affects PAK1 expression.
van der Klei
I J
IJ
Eukaryotic Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, NL-9751 NN Haren, The Netherlands. IJVDKLEI@BIOL.RUG.NL
van der Heide
M
M
Baerends
R J
RJ
Rechinger
K B
KB
Nicolay
K
K
Kiel
J A
JA
Veenhuis
M
M
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005656:Fungal Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol / Q000378:metabolism / Q000494:pharmacology; D008830:Microbodies / Q000737:chemistry / Q000235:genetics*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D009492:Neurospora crassa / Q000235:genetics; D016366:Open Reading Frames; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000235:genetics*; D010843:Pichia / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D017421:Sequence Analysis; D017385:Sequence Homology
D018076:DNA, Complementary; D005656:Fungal Proteins; C114262:PAK1 protein, Hansenula; D017853:Phosphotransferases (Alcohol Group Acceptor); C023132:glycerone kinase; D000432:Methanol