Mutually exclusive splicing of calcium-binding domain exons in chick alpha-actinin.
We have determined the complete sequence of chick brain alpha-actinin (892 amino acids; 107,644 Da). The sequence differs from that of smooth muscle alpha-actinin only in the region of the first EF-hand calcium-binding motif, where 27 residues in brain alpha-actinin are replaced by just 22 residues in the smooth muscle isoform. This probably accounts for the different calcium sensitivities of the two isoforms with respect to actin binding. Analysis of the gene structure showed that this region of sequence divergence is encoded by two separate exons whose incorporation is mutually exclusive. We have determined the proportion of the two transcripts in various tissues and cell lines using poly(A)+ RNA and a quantitative assay based on the polymerase chain reaction. MRC-5 fibroblasts and HeLa cells express mRNAs encoding both isoforms, whereas Namalwa lymphoblastoid cells, which lack actin stress fibers, express only the non-muscle mRNA. Both isoforms of alpha-actinin became incorporated into stress fibers and cell-matrix junctions when full-length chick alpha-actinin cDNAs were expressed in monkey COS cells. The levels of chick alpha-actinin mRNAs were found to be serum-inducible, suggesting that alpha-actinin may be an early response gene.
Waites
G T
GT
Department of Biochemistry, University of Leicester, United Kingdom.
Graham
I R
IR
Jackson
P
P
Millake
D B
DB
Patel
B
B
Blanchard
A D
AD
Weller
P A
PA
Eperon
I C
IC
Critchley
D R
DR
D000185:Actinin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D000199:Actins / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001921:Brain / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002645:Chickens; D005091:Exons; D005796:Genes; D005895:Gizzard, Avian / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000502:physiology; D009838:Oligodeoxyribonucleotides; D012326:RNA Splicing; D014162:Transfection
D000199:Actins; D009838:Oligodeoxyribonucleotides; D000185:Actinin; D002118:Calcium