Cell‐free Synthesis of a High Molecular Weight Relaxin‐related Protein*

Gast, Michael J.; Mercado-Simmen, Rosalia C.; Niall, Hugh D.; Boime, Irving

doi:10.1111/j.1749-6632.1980.tb47248.x

Cited by 14 publications

(6 citation statements)

References 18 publications

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“…In 1980, Gast et al (23) reported that RNA isolated from CL of pregnant sows directed the synthesis of a protein with immunological and sequence identity to authentic RLX. Later, evidence for synthesis of RLX was also found in translation mixtures using RNA isolated from CL of nonpregnant sows (24).…”

Section: Discussionmentioning

confidence: 99%

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Relaxin Gene Expression in the Sow Corpus Luteum during the Cycle, Pregnancy, and Lactation*

Bagnell

Tashima²,

Tsark³

et al. 1990

Endocrinology

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Relaxin (RLX) mRNA was sought in the corpus luteum of the pig during the cycle, pregnancy, and lactation using Northern analysis and in situ hybridization. Three oligonucleotide probes to regions of the preprorelaxin molecule were used for hybridization to detect RLX message and gave similar results. Northern analysis showed a single 1-kilobase RLX transcript at all three stages of the reproductive cycle studied. The intensity of the RLX hybridization signal was greatest in pregnancy and varied during the cycle. The signal in ovaries from day 3 cyclic animals increased by day 13 and declined on day 19. However, the hybridization signal at midcycle was only 2% of that during pregnancy. After parturition on day 2 of lactation, RLX message was still detected in the ovary, although at reduced levels compared with those during the cycle and pregnancy. In situ hybridization results showed hybridization to RLX mRNA in luteal tissue on day 13 of the cycle, but not on day 3 or 19. An increased hybridization signal was observed on days 40, 60, and 90 of pregnancy, with a decline on day 2 of lactation. Control sections incubated with labeled heterologous probe or preincubated with excess unlabeled probe did not hybridize. These results indicate a good correlation between the relative concentrations of RLX transcript and immunohistochemical results previously reported in the corpus luteum of the sow. In addition, they demonstrate that the RLX gene is expressed in luteal tissue, not only in pregnancy, but also in the cycle and early lactation.

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Section: Discussionmentioning

confidence: 99%

“…Two probes to adjacent regions of the C-peptide of porcine pro-RLX (C-peptide 1, AA 45-60; C-peptide 2, AA 61-76) were synthesized by Peninsula Laboratories (Belmont, CA). A third RLX-specific probe corresponding to a portion of the B-chain of pRLX (AA [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26] (12).…”

Section: Probesmentioning

confidence: 99%

Relaxin Gene Expression in the Sow Corpus Luteum during the Cycle, Pregnancy, and Lactation*

Bagnell

Tashima²,

Tsark³

et al. 1990

Endocrinology

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“…This suggests the existence of a signal sequence of approximately 3,000 daltons. Interestingly, evidence was obtained for mRNAs for relaxin in the nonpregnant ovary (54).…”

Section: Sites Of Productionmentioning

confidence: 94%

“…Its production by the follicle (65), the receptors for relaxin in the myoWinter, 1982 RELAXIN 85 metrium of the cyclic rat (25), the localization of mRNA for relaxin in the nonpregnant rat corpus luteum (31), the presence of mRNA in nonpregnant sow ovaries translatable into relaxin containing peptides in the wheat germ system (54), the detection of relaxin immunoactivity in cyclic rat ovarian extracts (170), the relaxin immunoactivity in cyclic guinea pig serum (158), the rises of relaxin immunoactivity on suckling in the sow (68), and the demonstrations of relaxin immunoactivity in human seminal plasma (81) all indicate that relaxin is a hormone of the nonpregnant female as well as of the male.…”

Section: Relaxin As a Local Hormonementioning

confidence: 99%

Relaxin as a New Hormone*

Bryant‐Greenwood

1982

Endocrine Reviews

137

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“…Rat preprorelaxin has been shown to contain a 105 amino acid connecting peptide linking the A and B chains (Hudson et al, 1981). Cell-free translations of mRNA isolated from the ovaries of pregnant pigs and immunoprecipitation of the protein products with relaxin antisera also suggested a large relaxin precursor of about 23,000 daltons as compared to 11,000 daltons for preproinsulin (Gast et al, 1980). It thus appeared likely that the pig relaxin precursor would be similar in structure to that of the rat with a large connecting peptide joining the A and B chains.…”

Section: Introductionmentioning

confidence: 95%

Porcine Relaxin: Molecular Cloning and cDNA Structure

Haley¹,

Hudson²,

Scanlon³

et al. 1982

DNA

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Relaxin is a peptide hormone produced by the corpora lutea of ovaries during pregnancy, softening and lengthening the ligaments of the pelvis and softening the cervix in order to make childbirth easier. In attempts to determine the nucleotide sequence coding for relaxin, recombinant DNA techniques were used to obtain a cDNA clone bank from total mRNA isolated from the ovaries of pigs in late pregnancy. Clones were screened using cDNA initiated by synthetic oligonucleotide primers coding for the Trp Val Glu Ile sequence of the porcine relaxin B chain. The synthetic undecamer [5'-ATCTCCACCCA-3'] was found to prime a specific 32P-labeled cDNA of approximately 300 nucleotides containing B chain and signal peptide coding sequences, as verified by nucleic acid sequence analysis. This cDNA was used to probe the ovarian clone bank. Several clones containing large inserts which hybridized to this probe were subjected to sequence analysis and some of these were found to contain the preprorelaxin coding region, comprising a signal peptide of 24 amino acids, a B chain of 32 amino acids, a large C peptide of 104 amino acids, and an A chain of 22 amino acids. From the amino acid sequence of prorelaxin derived in this way, it appears that the processing of prorelaxin involves two enzymes with chymotrypsin-like and trypsin-like specificity, respectively. In comparisons of porcine and rat preprorelaxins, the C region had as much amino acid sequence homology as the B and A chains. The C region is also rich in charged amino acids, suggesting a role for it beyond simply ensuring proper disulfide bond formation.

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Cell‐free Synthesis of a High Molecular Weight Relaxin‐related Protein*

Cited by 14 publications

References 18 publications

Relaxin Gene Expression in the Sow Corpus Luteum during the Cycle, Pregnancy, and Lactation*

Relaxin Gene Expression in the Sow Corpus Luteum during the Cycle, Pregnancy, and Lactation*

Relaxin as a New Hormone*

Porcine Relaxin: Molecular Cloning and cDNA Structure

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