Cloning and characterization of a glycogen synthase cDNA from human endometrium.
One major human uterine response to post-ovulatory progesterone is the accumulation of glycogen by the endometrium. A temporally related increase in glycogen synthase activity has been documented, but the isozyme responsible has not yet been identified. We have amplified a glycogen synthase (GS) complementary DNA (cDNA) from human endometrium by reverse transcription-polymerase chain reaction (RT-PCR). Overlapping clones of the PCR products provided a cDNA that is 3534 base pairs (bp) long, including a 22-bp poly(A)+ tail, and an open reading frame that encodes a 737 amino acid protein with a molecular weight of 83936. This cDNA is almost identical to that of human striated muscle GS. Differences include a double nucleotide substitution at 1983-1984 and five single nucleotide substitutions located, respectively, at positions 379, 2457, 2470, 2477, and 2553. These differences only alter the predicted amino acid sequence from that of the striated muscle protein by a single substitution at position 608. A 5'-end fragment plus an internal fragment of human myometrial GS cDNA were also analysed and were shown to have identity with the endometrial GS cDNA. Northern blot hybridization, using a human muscle-derived cDNA probe, detected the presence of a 4.0-kb GS messenger RNA (mRNA) in the endometrium and myometrium. Our results establish that the GS of human Mullerian tissues is, essentially, identical to that reported for human striated muscle.
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Department of Obstetrics and Gynecology, University of Wisconsin, Madison, U.S.A.
Schuler
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Shapiro
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004717:Endometrium / Q000201:enzymology*; D005260:Female; D006006:Glycogen Synthase / Q000235:genetics*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D006006:Glycogen Synthase