Determination of the prorelaxin nucleotide sequence and temporal expression of prorelaxin mRNA in the golden hamster / by R. Brian McCaslin.
| Author/creator | McCaslin, R. Brian author. |
| Other author | Renegar, Randall H., degree supervisor. |
| Other author | East Carolina University. Department of Biology. |
| Format | Theses and dissertations |
| Production | 1994. |
| Description | iv, 89 leaves : illustrations ; 28 cm |
| Supplemental Content | Access via ScholarShip |
| Subjects |
| Summary | Relaxin has been purified from the hamster placenta and the N-terminal amino acid sequence of this molecule has been determined. The objectives of this study were to determine the complete amino acid sequence of hamster prorelaxin and to evaluate the placental prorelaxin mRNA profile during gestation. Ovary, hippocampus and neocortex were also examined. A 21 bp oligonucleotide pool derived from the N-terminal amino acid sequence of hamster prorelaxin was used in 3' rapid amplification of cDNA ends methodology (3'RACE) to generate a specific cDNA for this molecule, and the nucleotide sequence was determined by the chain termination procedure. Total RNA was extracted from tissues using guanidinium hydrochloride methodology and the level of expression assessed by Northern analysis using the hamster prorelaxin cDNA. The deduced sequence for hamster prorelaxin contains 155 amino acids with 51.8, 42.9, and 38.2% homology to rat, human 1 and pig prorelaxin, respectively. Unique to hamster prorelaxin is an additional cysteine at position 9 of the B-chain. In addition this molecule contains multiple basic amino acids (RRRR) at the B-chain/C peptide cleavage site. Recent reports indicate that precursors containing multiple basic amino acids are cleaved by the enzyme furin in cells utilizing the constitutive secretory pathway. This information is consistent with previous studies from this laboratory suggesting that hamster relaxin is secreted by the constitutive pathway from placental trophoblast cells. Northern analysis of placental RNA identified a 900 bp transcript that was first detected on Day 10 of gestation. Intensity of the hybridization signal increased to the evening of Day 15 (parturition on Day 16) which paralleled previously determined tissue concentrations of relaxin immunoreactivity [Renegar, unpublished data]. Hybridization of the hamster prorelaxin cDNA to ovarian, hippocampal, or neocortical RNA was not detected following a 12 hour or 7 Day autoradiographic exposure. |
| General note | Submitted to the faculty of the Department of Biology. |
| General note | Advisor: Randall H. Renegar |
| Dissertation note | M.S. East Carolina University 1994 |
| Bibliography note | Includes bibliographical references (leaves 52-64). |
| Genre/form | Academic theses. |
| Genre/form | Academic theses. |
| Genre/form | Thèses et écrits académiques. |
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