Drosophila spectrin. II. Conserved features of the alpha-subunit are revealed by analysis of cDNA clones and fusion proteins.
Drosophila alpha-spectrin cDNA sequences were isolated from a lambda gt11 expression library. These cDNA clones encode fusion proteins that include portions of the Drosophila alpha-spectrin polypeptide as shown by a number of structural and functional criteria. The fusion proteins elicited antibodies that reacted strongly with Drosophila and vertebrate alpha-spectrins and a comparison of cyanogen bromide peptide maps demonstrated a clear structural correspondence between one fusion protein and purified Drosophila alpha-spectrin. Alpha-spectrin fusion protein also displayed calcium-dependent calmodulin-binding activity in blot overlay experiments and one fusion protein bound specifically to both Drosophila and bovine brain beta-spectrin subunits on protein blots. A region of the Drosophila cDNA cross-hybridized at lowered stringency with an avian alpha-spectrin cDNA. Together these data show that the composition, structure, and binding properties of the spectrin family of proteins have been remarkably well conserved between arthropods and vertebrates. Drosophila cDNA hybridized to an mRNA of greater than or equal to 9 kb on blots of total Drosophila poly A+ RNA; and hybridized in situ to a single site in polytene region 62B, 1-7. This result and Southern blot analysis of genomic DNA indicate that the sequences are likely to be single copy in the Drosophila genome.
Byers
T J
TJ
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
Dubreuil
R
R
Branton
D
D
Kiehart
D P
DP
Goldstein
L S
LS
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D004262:DNA Restriction Enzymes; D004330:Drosophila / Q000235:genetics*; D005796:Genes; D046911:Macromolecular Substances; D009693:Nucleic Acid Hybridization; D011994:Recombinant Proteins / Q000302:isolation & purification; D013049:Spectrin / Q000235:genetics* / Q000302:isolation & purification
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D013049:Spectrin; D004247:DNA; D004262:DNA Restriction Enzymes