Isolation and in Situ localization of a cDNA encoding a Kex2-like prohormone convertase in the nematode Caenorhabditis elegans

Eduardo Gómez-Saladín, David L. Wilson, Ian M. Dickerson

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

1. A cDNA that encodes a Kex2-like prohormone convertase (PC) containing an active site similar to that of mammalian PC2 has been isolated from C. elegans. Total RNA was isolated from a mixed population of strain BA713 worms. After poly-(A)-selection and reverse transcription, degenerate/nested polymerase chain reactions (PCR) were performed using primers based on conserved regions within the active sites of the known vertebrate and invertebrate endoproteases. 2. Two distinct 300-bp PCR products that shared homologies with the active sites of known Kex2-like endoproteases were isolated. These two PCR products were used to screen a C. elegans cDNA library. 3. The complete cDNA for a Kex2-like endoprotease, designated CELPC2, was isolated and determined to be 2527 bp in length. This size was confirmed by northern analysis. The deduced amino acid sequence for the CELPC2 cDNA is very similar to the known Kex2-like endoproteases, especially at conserved regions within the active sites, but not identical to any one of them. The strongest structural homology was to vertebrate and invertebrate PC2 sequences. 4. In situ hybridization suggests that CELPC2 is synthesized primarily in cells associated with the circumpharyngeal nerve ring and the dorsorectal ganglion.

Original languageEnglish
Pages (from-to)9-25
Number of pages17
JournalCellular and Molecular Neurobiology
Volume14
Issue number1
DOIs
StatePublished - Feb 1 1994

Fingerprint

Proprotein Convertases
Polymerase chain reaction
Caenorhabditis elegans
Catalytic Domain
Complementary DNA
Reaction products
Invertebrates
Polymerase Chain Reaction
Vertebrates
Poly A
Transcription
Gene Library
Ganglia
RNA
Reverse Transcription
In Situ Hybridization
Amino Acids
Amino Acid Sequence
Population

Keywords

  • Caenorhabditis elegans
  • endoprotease
  • Kex2
  • neuropeptide
  • PC2
  • prohormone convertase (PC)

ASJC Scopus subject areas

  • Neuroscience(all)
  • Genetics
  • Clinical Biochemistry
  • Cell Biology

Cite this

Isolation and in Situ localization of a cDNA encoding a Kex2-like prohormone convertase in the nematode Caenorhabditis elegans. / Gómez-Saladín, Eduardo; Wilson, David L.; Dickerson, Ian M.

In: Cellular and Molecular Neurobiology, Vol. 14, No. 1, 01.02.1994, p. 9-25.

Research output: Contribution to journalArticle

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