Class XIII myosins from the green alga Acetabularia: driving force in organelle transport and tip growth?
The green alga Acetabularia cliftonii (Dasycladales) contains at least two myosin genes, which already have been assigned class XIII of the myosin superfamily (Cope et al., 1996, Structure 4: 969-987). Here we report a complete analysis of their gene structure and their corresponding transcripts Aclmyo1 and Aclmyo2. Despite promising Northern blot data no evidence for alternative splicing could be found. Dissecting the primary structure at complementary deoxyribonucleic acid (cDNA) level we found a myosin typical organization in head, neck and variable tail region. Most striking is the extremely short tail region of Aclmyo1 with only 18 residues and the maximum number of 7 IQ motifs in Aclmyo2. Probing Acetabularia protein extracts with an antibody raised to a synthetic peptide derived from the amino terminal region in Alcmyo1 showed cross-reactivity to a polypeptide with a molecular mass of approximately 100 kD. This corresponds to the predicted molecular weight of Aclmyo1, which is 106 kD as deduced from the amino acid sequence. Additionally, the same cross-reactive protein is capable of binding F-actin as indicated by a co-sedimentation assay. Confocal laser scanning microscopy with raised antibody revealed co-localization with organelles, the budding region of lateral whorls and the cell apex suggesting involvement of putative Acetabularia myosin in organelle transport and tip growth.
Vugrek
Oliver
O
Max-Planck-Institute for Cell Biology, Rosenhof, 68526 Ladenburg, Germany. ovugrek@post.hinet.hr
Sawitzky
Heiko
H
Menzel
Diedrik
D
D020413:3' Untranslated Regions / Q000235:genetics; D000076:Acetabularia / Q000235:genetics* / Q000254:growth & development; D000199:Actins / Q000378:metabolism; D020418:Algal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D005091:Exons / Q000235:genetics; D005796:Genes / Q000235:genetics; D007438:Introns / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000502:physiology; D009218:Myosins / Q000235:genetics* / Q000378:metabolism; D015388:Organelles / Q000502:physiology; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D020413:3' Untranslated Regions; D000199:Actins; D020418:Algal Proteins; D012333:RNA, Messenger; D009218:Myosins