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  • Author: G W Tregear x
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Structure of rhesus monkey relaxin predicted by analysis of the single-copy rhesus monkey relaxin gene

R. J. Crawford, V. E. Hammond, P. J. Roche, P. D. Johnston, and G. W. Tregear

ABSTRACT

The gene encoding rhesus monkey relaxin has been investigated. A cDNA library was prepared using corpus luteal RNA from a pregnant rhesus monkey, cDNA clones encoding relaxin were isolated and the nucleotide sequence was determined. The amino acid sequence of rhesus monkey preprorelaxin, predicted from the cDNA, demonstrates that the sequence has not been strongly conserved when compared with that of man, although features characteristic of the relaxin molecule have been maintained. This structural information will allow production of rhesus monkey relaxin, leading to studies investigating the bioactivity of relaxin in a homologous primate system. Southern blot analysis indicated that there is only one relaxin gene in the rhesus monkey and baboon genomes. In this respect these primate genomes are different from the human genome which contains two relaxin genes.

DOI:
https://doi.org/10.1677/jme.0.0030169
Volume 3: Issue 3,
Pages:
169–174 (6 total)
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Characterization of human relaxin gene regulation in the relaxin-expressing human prostate adenocarcinoma cell line LNCaP.FGC

J M Gunnersen, P J Roche, G W Tregear, and R J Crawford

ABSTRACT

Relaxin is a peptide hormone which is produced in human reproductive tissues including the ovary and prostate gland. Little is known of the molecular events regulating relaxin gene transcription. We have studied this question using gene transfer of relaxin promoter/reporter gene constructs into a relaxin-expressing cell line. A number of human cell lines expressed relaxin as detected by reverse transcription-PCR. In one of these lines, the prostate adenocarcinoma cell line LNCaP.FGC, relaxin mRNA was also detected by Northern blot analysis. The DNA sequences of the proximal 5′-flanking regions (∼900 nucleotides) of the two human relaxin genes, HI and H2, were determined. Deletion constructs containing portions of the 5′-flanking regions of HI and H2 linked to the bacterial chloramphenicol acetyl transferase reporter gene were prepared. The expression of the reporter gene constructs was analysed in the LNCaP.FGC cell line and the results of these transient transfection assays have led to the identification of positive and negative regulatory regions within the 5′-flanking DNA. A difference in activity of the H1 and H2 gene promoters in this prostate cell line was observed, with the H2 promoter being more active. This situation may mimic that occurring in vivo since the relaxin secreted from the prostate gland into seminal fluid is the product of the H2 gene.

DOI:
https://doi.org/10.1677/jme.0.0150153
Volume 15: Issue 2,
Pages:
153–166 (14 total)
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The mouse relaxin gene: nucleotide sequence and expression

B A Evans, M John, K J Fowler, R J Summers, M Cronk, J Shine, and G W Tregear

ABSTRACT

Relaxin is a polypeptide hormone that has a variety of physiological effects both on remodelling of collagen and on uterine contractility. These are most apparent during pregnancy. The sequences of relaxin cDNAs derived from ovaries of late-pregnant random-bred Swiss mice have been established. Multiple subclones obtained from three independent polymerase chain reaction experiments were found to encode relaxins which were identical except at position 11 in the A chain (Ile or Val). All mouse relaxin cDNAs expressed in the ovary during pregnancy had an extra tyrosine inserted prior to the final A chain cysteine residue, a result confirmed by direct sequencing of relaxin peptides. Whilst this tyrosine insertion must have local effects on the folding of the A chain, structure—activity studies will clarify whether it perturbs functional interaction with the relaxin receptor. We have shown that there is a single relaxin gene in the mouse genome, and that expression during pregnancy occurs in the ovary but is not detectable in the placenta, uterus or fetus.

DOI:
https://doi.org/10.1677/jme.0.0100015
Volume 10: Issue 1,
Pages:
15–23 (9 total)