Molecular cloning of macrophin, a human homologue of Drosophila kakapo with a close structural similarity to plectin and dystrophin.
We have determined the complete cDNA coding sequence of a novel cytoskeletal protein by the degenerative primer-mediated PCR strategy. This novel gene, named as macrophin (microfilament and actin filament cross-linker protein related to plectin and dystrophin, Accession No. AB029290), appears to be a human homologue of a Drosophila gene, kakapo, and shows close similarity to plectin and dystrophin on the search of BLAST homology-computed database. Comparison of the deduced protein sequences for macrophin and kakapo revealed that they were 66% similar, and both of them have an NH2-terminal actin-binding domain, a central rod region composed of spectrin-like repeats, and COOH-terminal Gas2-related region. The predicted sequences for macrophin are 5430 amino acids in length with a calculated molecular mass of 620 kDa, which is one of the largest size idenfied in human cytoskeletal proteins. High expression of macrophin was observed in brain, heart, lung, placenta, liver, kidney, and pancreas. In pancreas, we found that macrophin was specifically expressed in acinar cells than islet cells by in situ hybridization. By using radiation hybrid panel, we have mapped the macrophin gene to the chromosome 1p31-32.
Okuda
T
T
Department of Radiology, Department of Molecular Pathogenesis, Pathology II, Nagoya University School of Medicine, 65 Turumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
Matsuda
S
S
Nakatsugawa
S
S
Ichigotani
Y
Y
Iwahashi
N
N
Takahashi
M
M
Ishigaki
T
T
Hamaguchi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002874:Chromosome Mapping; D002878:Chromosomes, Human, Pair 1; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D004330:Drosophila / Q000235:genetics; D029721:Drosophila Proteins; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics; D007381:Intermediate Filament Proteins / Q000737:chemistry / Q000235:genetics*; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010179:Pancreas / Q000378:metabolism; D051190:Plectin; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid
D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D029721:Drosophila Proteins; D016189:Dystrophin; D019476:Insect Proteins; D007381:Intermediate Filament Proteins; D008840:Microfilament Proteins; C495332:PLEC protein, human; D051190:Plectin; D012333:RNA, Messenger; C450828:shot protein, Drosophila