Molecular cloning, cDNA sequence, and localization of a prohormone convertase (PC2) from the Aplysia atrial gland

G. T. Nagle, A. T. Garcia, S. L. Knock, E. L. Gorham, W. R A Van Heumen, A. Kurosky

    Research output: Contribution to journalArticle

    19 Citations (Scopus)

    Abstract

    Neuropeptides and peptide hormones are synthesized as part of larger precursor proteins that are processed post-translationally by subtilisin- related calcium-dependent prohormone convertases (PCs), frequently at multiple basic sites, to generate biologically active peptides. The atrial gland of Aplysia californica produces large quantities of egg-laying hormone (ELH)-related peptides, providing a unique opportunity to study prohormone processing. We have screened an Aplysia atrial gland cDNA library using a Lymnaea stagnalis PC2 probe and have isolated an Aplysia PC2-related 4.6-kb cDNA partial clone that was truncated on the 5' end. The remaining 5' atrial gland PC2 nucleotide sequence was obtained by reverse transcription/polymerase chain reaction (RT-PCR). The composite cDNA structure (5.6 kb) was deduced from sequence analysis of the RT-PCR product combined with the sequence obtained from the cDNA clone. The deduced cDNA of Aplysia atrial gland PC2 encoded a putative preproendoprotease of 653 amino acids that was evolutionarily related to other eukaryotic PC2s, and showed the strongest sequence identity with recently reported Aplysia nervous tissue PC2 sequences. In situ hybridization demonstrated extensive expression of PC2 in atrial gland secretory cells. The cDNA clone contained a relatively long 3' untranslated region (3'-UTR) of 3,632 nucleotides. Strikingly, the 3'-UTR also contained several major nucleotide repeat sequences including the microsatellite repeats, (CA)(n) and (TG)(n), and a TA-rich region comprised largely of the triplet repeat (TTA)(n). The characterized Aplysia PC2 is a candidate endoprotease that may play an important role in the processing of ELH-related precursors in the atrial gland and represents the first example of PC2 expression in exocrine tissue.

    Original languageEnglish (US)
    Pages (from-to)145-154
    Number of pages10
    JournalDNA and Cell Biology
    Volume14
    Issue number2
    StatePublished - 1995

    Fingerprint

    Proprotein Convertase 2
    Aplysia
    Molecular Cloning
    Complementary DNA
    Peptide Hormones
    Clone Cells
    3' Untranslated Regions
    Reverse Transcription
    Ovum
    Proprotein Convertases
    Lymnaea
    Trinucleotide Repeats
    Subtilisin
    Nerve Tissue
    Polymerase Chain Reaction
    Protein Precursors
    Neuropeptides
    Gene Library
    Microsatellite Repeats
    In Situ Hybridization

    ASJC Scopus subject areas

    • Cell Biology
    • Genetics
    • Molecular Biology

    Cite this

    Nagle, G. T., Garcia, A. T., Knock, S. L., Gorham, E. L., Van Heumen, W. R. A., & Kurosky, A. (1995). Molecular cloning, cDNA sequence, and localization of a prohormone convertase (PC2) from the Aplysia atrial gland. DNA and Cell Biology, 14(2), 145-154.

    Molecular cloning, cDNA sequence, and localization of a prohormone convertase (PC2) from the Aplysia atrial gland. / Nagle, G. T.; Garcia, A. T.; Knock, S. L.; Gorham, E. L.; Van Heumen, W. R A; Kurosky, A.

    In: DNA and Cell Biology, Vol. 14, No. 2, 1995, p. 145-154.

    Research output: Contribution to journalArticle

    Nagle, GT, Garcia, AT, Knock, SL, Gorham, EL, Van Heumen, WRA & Kurosky, A 1995, 'Molecular cloning, cDNA sequence, and localization of a prohormone convertase (PC2) from the Aplysia atrial gland', DNA and Cell Biology, vol. 14, no. 2, pp. 145-154.
    Nagle GT, Garcia AT, Knock SL, Gorham EL, Van Heumen WRA, Kurosky A. Molecular cloning, cDNA sequence, and localization of a prohormone convertase (PC2) from the Aplysia atrial gland. DNA and Cell Biology. 1995;14(2):145-154.
    Nagle, G. T. ; Garcia, A. T. ; Knock, S. L. ; Gorham, E. L. ; Van Heumen, W. R A ; Kurosky, A. / Molecular cloning, cDNA sequence, and localization of a prohormone convertase (PC2) from the Aplysia atrial gland. In: DNA and Cell Biology. 1995 ; Vol. 14, No. 2. pp. 145-154.
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