Studies on the expression and evolution of the glycogen phosphorylase gene family in the rat.
Muscle, liver, and brain glycogen phosphorylases in mammals comprise a family of closely related isozymes that are differentially expressed in a wide variety of cell types. Towards obtaining a better understanding of the mechanisms governing the tissue-specific control of expression of this isozyme family, we used an antibody generated against bovine liver phosphorylase to obtain quantitative estimates of the concentrations of the three isozymes in rat tissues by Western blot analysis. This analysis indicated that expression of these isozymes at the protein level, although widespread, was tissue-specific and each isozyme exhibited variations in expression throughout the tissues where it was produced. We also began a preliminary analysis of the evolution of the genes encoding these three isozymes. Towards this end, we isolated and sequenced a partial cDNA to the rat brain isozyme that encompassed the coding region from amino acids 569 to 729. Using known phosphorylase gene sequences, we reconstructed a phylogeny spanning three kingdoms. This phylogeny indicated that brain and muscle isozymes are more closely related to each other than to the liver isozyme and that gene duplications that give rise to the family predate the mammalian radiation. Differences in the relative rates of change of the three isozymes were observed and this may reflect different constraints on their evolution perhaps related to their functional roles and (or) tissue-specific expression.
Crerar
M M
MM
Department of Biology, York University, North York, Ont., Canada.
Hudson
J W
JW
Matthews
K E
KE
David
E S
ES
Golding
G B
GB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D015152:Blotting, Northern; D015153:Blotting, Western; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005796:Genes; D007527:Isoenzymes / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity; D006005:Phosphorylases / Q000235:genetics*; D051381:Rats; D011919:Rats, Inbred Strains / Q000235:genetics*; D014158:Transcription, Genetic
D007527:Isoenzymes; D004247:DNA; D006005:Phosphorylases