Structural characterization of the rat carboxypeptidase-E gene

Yong Keun Jung, Cheryl J. Kunczt, Randy K. Pearson, Jack E. Dixon, Lloyd D. Fricker

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Several genomic clones encoding carboxypeptidase-E (CPE) have been isolated and partially sequenced. Southern blot analysis indicates that a single copy of this gene is present in the rat genome. The entire gene spans approximately 50 kilobases and consists of nine exons, each of which contains protein-coding regions. Only one of the exon/intron junctions of the rat CPE gene is present in a comparable position within the genes for carboxypeptidase-A and -B, both of which are only 17-21% homologous to CPE at the amino acid level. Nuclease protection analysis shows that alternative splicing of exons 7, 8, and 9 does not occur, indicating that the heterogeneity of the C-terminal region of CPE is due to posttranslational processing. Primer extension and nuclease protection analyses have identified the 5′ end of CPE mRNA to be 105 nucleotides up-stream from the ATG used for protein translation. The 5′ flanking region does not contain TATA and/or CCAAT boxes in the near vicinity of the transcription initiation site. The 5′ flanking region is GC rich, containing 70% GC residues over nucleotides -1 to -150 (relative to the transcription initiation site). Putative consensus sites for the enhancer elements SP-1, NF-1, Pan-1, and AP-2 are present in the region from -60 to -330. Since this report describes the first neuropeptide-processing enzyme gene to be partially sequenced, it is not possible to compare the sequence with those of other processing enzymes that show similar tissue-specific expression. However, comparison of the CPE sequence with 5′flanking regions of other neuroendocrine genes has revealed a short region (12-18 nucleotides) that is highly conserved among CPE, neuropeptide-Y, oxytocin, insulin, and tyrosine hy-droxylase genes.

Original languageEnglish (US)
Pages (from-to)1257-1268
Number of pages12
JournalMolecular Endocrinology
Volume5
Issue number9
StatePublished - 1991

Fingerprint

Carboxypeptidase H
Genes
Exons
Nucleotides
5' Flanking Region
Transcription Initiation Site
Carboxypeptidase B
Carboxypeptidases A
Gene Order
Neuropeptide Y
Transcription Factor AP-1
Alternative Splicing
Protein Biosynthesis
Oxytocin
Enzymes
Southern Blotting
Neuropeptides
Introns
Open Reading Frames
Tyrosine

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Jung, Y. K., Kunczt, C. J., Pearson, R. K., Dixon, J. E., & Fricker, L. D. (1991). Structural characterization of the rat carboxypeptidase-E gene. Molecular Endocrinology, 5(9), 1257-1268.

Structural characterization of the rat carboxypeptidase-E gene. / Jung, Yong Keun; Kunczt, Cheryl J.; Pearson, Randy K.; Dixon, Jack E.; Fricker, Lloyd D.

In: Molecular Endocrinology, Vol. 5, No. 9, 1991, p. 1257-1268.

Research output: Contribution to journalArticle

Jung, YK, Kunczt, CJ, Pearson, RK, Dixon, JE & Fricker, LD 1991, 'Structural characterization of the rat carboxypeptidase-E gene', Molecular Endocrinology, vol. 5, no. 9, pp. 1257-1268.
Jung YK, Kunczt CJ, Pearson RK, Dixon JE, Fricker LD. Structural characterization of the rat carboxypeptidase-E gene. Molecular Endocrinology. 1991;5(9):1257-1268.
Jung, Yong Keun ; Kunczt, Cheryl J. ; Pearson, Randy K. ; Dixon, Jack E. ; Fricker, Lloyd D. / Structural characterization of the rat carboxypeptidase-E gene. In: Molecular Endocrinology. 1991 ; Vol. 5, No. 9. pp. 1257-1268.
@article{e39afc6de497401881759c033fc14b7f,
title = "Structural characterization of the rat carboxypeptidase-E gene",
abstract = "Several genomic clones encoding carboxypeptidase-E (CPE) have been isolated and partially sequenced. Southern blot analysis indicates that a single copy of this gene is present in the rat genome. The entire gene spans approximately 50 kilobases and consists of nine exons, each of which contains protein-coding regions. Only one of the exon/intron junctions of the rat CPE gene is present in a comparable position within the genes for carboxypeptidase-A and -B, both of which are only 17-21{\%} homologous to CPE at the amino acid level. Nuclease protection analysis shows that alternative splicing of exons 7, 8, and 9 does not occur, indicating that the heterogeneity of the C-terminal region of CPE is due to posttranslational processing. Primer extension and nuclease protection analyses have identified the 5′ end of CPE mRNA to be 105 nucleotides up-stream from the ATG used for protein translation. The 5′ flanking region does not contain TATA and/or CCAAT boxes in the near vicinity of the transcription initiation site. The 5′ flanking region is GC rich, containing 70{\%} GC residues over nucleotides -1 to -150 (relative to the transcription initiation site). Putative consensus sites for the enhancer elements SP-1, NF-1, Pan-1, and AP-2 are present in the region from -60 to -330. Since this report describes the first neuropeptide-processing enzyme gene to be partially sequenced, it is not possible to compare the sequence with those of other processing enzymes that show similar tissue-specific expression. However, comparison of the CPE sequence with 5′flanking regions of other neuroendocrine genes has revealed a short region (12-18 nucleotides) that is highly conserved among CPE, neuropeptide-Y, oxytocin, insulin, and tyrosine hy-droxylase genes.",
author = "Jung, {Yong Keun} and Kunczt, {Cheryl J.} and Pearson, {Randy K.} and Dixon, {Jack E.} and Fricker, {Lloyd D.}",
year = "1991",
language = "English (US)",
volume = "5",
pages = "1257--1268",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "9",

}

TY - JOUR

T1 - Structural characterization of the rat carboxypeptidase-E gene

AU - Jung, Yong Keun

AU - Kunczt, Cheryl J.

AU - Pearson, Randy K.

AU - Dixon, Jack E.

AU - Fricker, Lloyd D.

PY - 1991

Y1 - 1991

N2 - Several genomic clones encoding carboxypeptidase-E (CPE) have been isolated and partially sequenced. Southern blot analysis indicates that a single copy of this gene is present in the rat genome. The entire gene spans approximately 50 kilobases and consists of nine exons, each of which contains protein-coding regions. Only one of the exon/intron junctions of the rat CPE gene is present in a comparable position within the genes for carboxypeptidase-A and -B, both of which are only 17-21% homologous to CPE at the amino acid level. Nuclease protection analysis shows that alternative splicing of exons 7, 8, and 9 does not occur, indicating that the heterogeneity of the C-terminal region of CPE is due to posttranslational processing. Primer extension and nuclease protection analyses have identified the 5′ end of CPE mRNA to be 105 nucleotides up-stream from the ATG used for protein translation. The 5′ flanking region does not contain TATA and/or CCAAT boxes in the near vicinity of the transcription initiation site. The 5′ flanking region is GC rich, containing 70% GC residues over nucleotides -1 to -150 (relative to the transcription initiation site). Putative consensus sites for the enhancer elements SP-1, NF-1, Pan-1, and AP-2 are present in the region from -60 to -330. Since this report describes the first neuropeptide-processing enzyme gene to be partially sequenced, it is not possible to compare the sequence with those of other processing enzymes that show similar tissue-specific expression. However, comparison of the CPE sequence with 5′flanking regions of other neuroendocrine genes has revealed a short region (12-18 nucleotides) that is highly conserved among CPE, neuropeptide-Y, oxytocin, insulin, and tyrosine hy-droxylase genes.

AB - Several genomic clones encoding carboxypeptidase-E (CPE) have been isolated and partially sequenced. Southern blot analysis indicates that a single copy of this gene is present in the rat genome. The entire gene spans approximately 50 kilobases and consists of nine exons, each of which contains protein-coding regions. Only one of the exon/intron junctions of the rat CPE gene is present in a comparable position within the genes for carboxypeptidase-A and -B, both of which are only 17-21% homologous to CPE at the amino acid level. Nuclease protection analysis shows that alternative splicing of exons 7, 8, and 9 does not occur, indicating that the heterogeneity of the C-terminal region of CPE is due to posttranslational processing. Primer extension and nuclease protection analyses have identified the 5′ end of CPE mRNA to be 105 nucleotides up-stream from the ATG used for protein translation. The 5′ flanking region does not contain TATA and/or CCAAT boxes in the near vicinity of the transcription initiation site. The 5′ flanking region is GC rich, containing 70% GC residues over nucleotides -1 to -150 (relative to the transcription initiation site). Putative consensus sites for the enhancer elements SP-1, NF-1, Pan-1, and AP-2 are present in the region from -60 to -330. Since this report describes the first neuropeptide-processing enzyme gene to be partially sequenced, it is not possible to compare the sequence with those of other processing enzymes that show similar tissue-specific expression. However, comparison of the CPE sequence with 5′flanking regions of other neuroendocrine genes has revealed a short region (12-18 nucleotides) that is highly conserved among CPE, neuropeptide-Y, oxytocin, insulin, and tyrosine hy-droxylase genes.

UR - http://www.scopus.com/inward/record.url?scp=0026394542&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026394542&partnerID=8YFLogxK

M3 - Article

VL - 5

SP - 1257

EP - 1268

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 9

ER -