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.
Eukaryotic Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, NL-9751 NN Haren, The Netherlands. IJVDKLEI@BIOL.RUG.NL
I J
IJ
van der Klei
M
M
van der Heide
R J
RJ
Baerends
K B
KB
Rechinger
K
K
Nicolay
J A
JA
Kiel
M
M
Veenhuis
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