Molecular cloning of the myelin basic proteins in the shark, Squalus acanthias, and the ray, Raja erinacia.
Myelin basic proteins (MBPs) are a family of alternatively spliced isoforms present in myelin sheaths of most vertebrates. A reverse transcriptase-polymerase chain reaction (RT-PCR) approach was used to clone MBP isoforms in species representing two superorders of elasmobranchs: Squalus acanthias, representing Squalomorph sharks, and Raja erinacia, representing Batoidea rays. Two products were generated from each species. The larger product encoded a 155 amino acid protein, the same size as MBPs from two Galeomorph sharks, Heterodontus francisci and Carcharhinus obscurus, which, based upon alignment with other vertebrate MBPs, contained six of the seven MBP exons; only exon II was absent. The smaller product encoded a 141 amino acid protein that lacked exon II and exon V. There were 26 and 30 nucleotide differences between Squalus and Heterodontus, and Raja and Heterodontus, respectively. Sequences from Squalus and Raja were far more similar, having only five nucleotide differences. Both isoforms of elasmobranch MBP contain 18.5% basic (lysine plus arginine) amino acids, compared with 17.5% in mammalian MBPs comprised of the corresponding exons. Northern blot analysis of whole brain total RNA revealed a single band of 2.5 kb in Squalus, and three bands of 1.2, 1.4, and 2.3 kb in Raja. The finding that MBPs of a Squalomorph shark and a Batoidea ray are closer to one another than either is to the Galeomorph sharks suggests that MBP sequence information may prove useful in classifying modern day Chondrichthytes.
Spivack
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WD
Marine Biological Laboratory, Woods Hole, Massachusetts.
Zhong
N
N
Salerno
S
S
Saavedra
R A
RA
Gould
R M
RM
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle; D002645:Chickens; D003001:Cloning, Molecular; D005091:Exons; D006801:Humans; D007536:Isomerism; D051379:Mice; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D012194:RNA-Directed DNA Polymerase; D011817:Rabbits; D051381:Rats; D012754:Sharks / Q000378:metabolism*; D012860:Skates, Fish / Q000378:metabolism*
D004676:Myelin Basic Protein; D012313:RNA; D012194:RNA-Directed DNA Polymerase