Cloning and sequencing of cDNA and genomic DNA encoding PDM phosphatase of Fusarium moniliforme.
PDM phosphatase was purified approximately 500-fold through six steps from the extract of dried powder of the culture filtrate of Fusarium moniliforme. The purified preparation appeared homogeneous on SDS-PAGE although the protein band was broad. Amino acid sequence information was collected on tryptic peptides from this preparation. cDNA cloning was carried out based on the information. A full-length cDNA was obtained and sequenced. The sequence had an open reading frame of 651 amino acid residues with a molecular mass of 69,988 Da. Cloning and sequencing of the genomic DNA corresponding to the cDNA was also conducted. The deduced amino acid sequence could account for many but not all of the tryptic peptides, suggesting presence of contaminant protein(s). SDS-PAGE analysis after chemical deglycosylation showed two proteins with molecular masses of 58 and 68 kDa. This implied that the 58 kDa protein had been copurified with PDM phosphatase. Homology search showed that PDM phosphatase belongs to the purple acid phosphatase family, which is widely distributed in the biosphere. Sequence data of fungal purple acid phosphatases were collected from the database. Processing of the data revealed presence of two types, whose evolutionary relationships were discussed.
Yoshida
Hiroshi
H
Department of Chemistry, Faculty of Medicine, Shimane University, 89-1 En-ya-cho, Izumo 693-8501. yoshidah@withe.ne.jp
Iizuka
Mari
M
Narita
Takao
T
Norioka
Naoko
N
Norioka
Shigemi
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry*; D004271:DNA, Fungal / Q000737:chemistry*; D005656:Fungal Proteins; D005670:Fusarium / Q000201:enzymology; D008969:Molecular Sequence Data; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D016415:Sequence Alignment
D018076:DNA, Complementary; D004271:DNA, Fungal; D005656:Fungal Proteins; D010744:Phosphoric Monoester Hydrolases; C019047:phosphodiesterase-phosphomonoesterase; D010727:Phosphoric Diester Hydrolases