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Cloning of two novel growth hormone transcripts expressed in human placenta.

Journal article
Authors C L Boguszewski
Per-Arne Svensson
T Jansson
R Clark
L M Carlsson
B Carlsson
Published in The Journal of clinical endocrinology and metabolism
Volume 83
Issue 8
Pages 2878-85
ISSN 0021-972X
Publication year 1998
Published at Institute of Internal Medicine, Dept of Medicine
Pages 2878-85
Language en
Links dx.doi.org/10.1210/jcem.83.8.5017
www.ncbi.nlm.nih.gov/entrez/query.f...
Keywords Alternative Splicing, Amino Acid Sequence, Base Sequence, Chemical Phenomena, Chemistry, Physical, Cloning, Molecular, DNA, Complementary, Female, Gene Expression, Genetic Variation, Human Growth Hormone, chemistry, genetics, Humans, Molecular Sequence Data, Placenta, chemistry, Polymerase Chain Reaction, Pregnancy, RNA, Messenger, analysis, genetics, RNA-Directed DNA Polymerase, Sequence Analysis, DNA
Subject categories Basic Medicine

Abstract

Several isoforms of human GH (hGH) are produced by two related genes expressed in the pituitary (hGH-N) and in the placenta (hGH-V). These genes consist of five exons (denoted 1-5) separated by four introns (denoted A-D). In the present report, two new transcripts of the hGH-V gene are described. The coding region of the hGH-V gene was amplified by RT-PCR using placental complementary DNA as template. DNA sequencing of several clones revealed two novel transcripts. One had a 45-bp deletion caused by the use of an alternative splice acceptor site within exon 3, similar to that in the hGH-N gene, predicting a 20-kDa isoform of hGH-V. The other transcript was generated by the use of an alternative splice donor site causing a 4-bp deletion in the end of exon 4, predicting a 24-kDa protein with 219 amino acids, which we refer to as hGH-V3. The carboxy-terminal sequence of hGH-V3 differs from 22-kDa hGH-V and hGH-V2, the two previously reported transcripts of the hGH-V gene, and does not contain a predicted transmembrane domain as described for hGH-V2. Ligase chain reaction was then used to analyze the possible use of the same splicing pattern in transcripts derived from the other genes of the hGH-gene cluster. Alternatively spliced transcripts encoding the 20-kDa hGH isoform were detected from the hGH-N and hGH-V genes, but not from the human chorionic somatomammotropin-A/B genes. The alternative splicing generating hGH-V3 was only demonstrated in transcripts derived from the hGH-V gene. Using competitive RT-PCR, the expression of hGH-V3 was estimated to be 10% of the hGH-V messenger RNA in full-term normal placentas and in placentas from pathological pregnancies. The 20-kDa hGH-V was detected in two of four full-term normal placentas, whereas a weak signal was observed in one of the pathological placentas. We conclude that the hGH-V primary transcript undergoes alternative splicing pathways generating at least four different messenger RNAs, predicting the expression of different hGH isoforms, including two with a complete sequence divergence in the carboxy-terminus.

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