Molecular cloning and characterization of human trabeculin-alpha, a giant protein defining a new family of actin-binding proteins.
We describe the molecular cloning and characterization of a novel giant human cytoplasmic protein, trabeculin-alpha (M(r) = 614,000). Analysis of the deduced amino acid sequence reveals homologies with several putative functional domains, including a pair of alpha-actinin-like actin binding domains; regions of homology to plakins at either end of the giant polypeptide; 29 copies of a spectrin-like motif in the central region of the protein; two potential Ca(2+)-binding EF-hand motifs; and a Ser-rich region containing a repeated GSRX motif. With similarities to both plakins and spectrins, trabeculin-alpha appears to have evolved as a hybrid of these two families of proteins. The functionality of the actin binding domains located near the N terminus was confirmed with an F-actin binding assay using glutathione S-transferase fusion proteins comprising amino acids 9-486 of the deduced peptide. Northern and Western blotting and immunofluorescence studies suggest that trabeculin is ubiquitously expressed and is distributed throughout the cytoplasm, though the protein was found to be greatly up-regulated upon differentiation of myoblasts into myotubes. Finally, the presence of cDNAs similar to, yet distinct from, trabeculin-alpha in both human and mouse suggests that trabeculins may form a new subfamily of giant actin-binding/cytoskeletal cross-linking proteins.
Sun
Y
Y
Dana Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
Zhang
J
J
Kraeft
S K
SK
Auclair
D
D
Chang
M S
MS
Liu
Y
Y
Sutherland
R
R
Salgia
R
R
Griffin
J D
JD
Ferland
L H
LH
Chen
L B
LB
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D015638:Cytochalasin D / Q000494:pharmacology; D006801:Humans; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D015739:Nocodazole / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid
C095003:MACF1 protein, human; C490632:Macf1 protein, mouse; D008840:Microfilament Proteins; D015638:Cytochalasin D; D015739:Nocodazole