Molecular characterization of glycogen synthase 1 and its tissue expression profile with type II hexokinase and muscle-type phosphofructokinase in horses.
Muscle glycogen synthase (GYS1) is the rate-limiting enzyme in glycogen synthesis, and its activity is regulated by the phosphorylation states of certain amino acid residues encoded by the GYS1 gene. In the present study, the authors molecularly characterized the full-length equine GYS1 (eGYS1) cDNA and found that it contains a less common polyadenylation signal (AATACA). An amino acid alignment with other mammalian GYS1 showed that the phosphorylation sites in eGYS1 are completely conserved. Genomic DNA analysis revealed that the equine-specific substitutions (Glu 16 Asp and Ala 252 Thr) were completely conserved among six equine species. The tissue expression profiles of eGYS1, equine type II hexokinase (eHKII) and muscle-type phosphofructokinase (ePFKM) were determined by real-time PCR and western blot analysis. The mRNA expression level of eGYS1 was significantly higher in the cervical muscle as compared to other tissues. The cervical muscle and heart tissue samples contained a broad range of eGYS1 protein bands that appeared to reflect multiple phosphorylation states. eHKII was predominately expressed only in the cervical muscle; unlike its expression in other mammals, eHKII was not substantially expressed in the insulin-responsive heart or adipose tissue of horse. The expression level of ePFKM mRNA was significantly higher in the heart than in the cervical muscle, which differs from the PFKM expression pattern of other mammals. These tissue expression profiles are fundamental for the understanding of equine glucose metabolism.
Echigoya
Yusuke
Y
Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa, 252-8510, Japan.
Okabe
Hirotarou
H
Itou
Takuya
T
Endo
Hideki
H
Sakai
Takeo
T
D000818:Animals; D015153:Blotting, Western; D001947:Breeding; D005091:Exons / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016678:Genome / Q000235:genetics; D006006:Glycogen Synthase / Q000235:genetics* / Q000378:metabolism; D006593:Hexokinase / Q000235:genetics* / Q000378:metabolism; D006736:Horses / Q000235:genetics*; D009928:Organ Specificity / Q000235:genetics; D025203:Phosphofructokinase-1, Muscle Type / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D012333:RNA, Messenger; D006006:Glycogen Synthase; D025203:Phosphofructokinase-1, Muscle Type; D006593:Hexokinase
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