Structure and expression of the cAMP cell-surface receptor

C. L. Saxe, P. Klein, T. J. Sun, A. R. Kimmel, Peter N Devreotes

Research output: Contribution to journalArticle

Abstract

Using antibodies specific for the 3',5'-cyclic AMP (cAMP) cell surface receptor of Dictyostelium discoideum, we have screened λgtll expression libraries and isolated a series of cDNAs derived from cAMP receptor mRNA during early development. The identity of the cDNA clones was verified by multiple criteria: 1) β-galactosidase fusion proteins synthesized by isolated cDNA clones stain intensely with cAMP receptor directed antiserum, 2) these fusion proteins affinity purify antibodies specific for the cAMP receptor, 3) the cDNA probes hybridize to a 2 kb mRNA whose change in relative level of abundance during development parallels that of receptor mRNA as assayed by in vitro translation, 4) the 2 kb mRNA size equals that of receptor mRNA as determined by in vitro translation of size fractionated poly (A)+ RNA, and 5) RNA transcribed in vitro from cDNAs containing the entire protein-coding region produces a polypeptide by in vitro translation with an apparent molecular weight in close agreement with that of nascent cAMP receptor protein produced by in vitro translation of cellular RNA. The DNA sequence predicts an open reading frame of 392 amino acids. The deduced amino acid sequence contains seven domains enriched in hydrophobic residues. A model is proposed in which the cAMP cell-surface receptor traverses the lipid bilayer seven times in a pattern similar to that of other receptors, such as rhodopsin, which interact with G-proteins. The structural similarities suggest a gene family of related surface receptors from such evolutionarily diverse species as Dictyostelium, yeast, and mammals.

Original languageEnglish (US)
Pages (from-to)227-235
Number of pages9
JournalDevelopmental Genetics
Volume9
Issue number4-5
StatePublished - 1988

Fingerprint

Cell Surface Receptors
Cyclic AMP
Cyclic AMP Receptors
Complementary DNA
Messenger RNA
Dictyostelium
Open Reading Frames
Clone Cells
Cyclic AMP Receptor Protein
Galactosidases
RNA
Antibody Affinity
Rhodopsin
Lipid Bilayers
GTP-Binding Proteins
Immune Sera
Mammals
Amino Acid Sequence
Proteins
Coloring Agents

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Genetics

Cite this

Saxe, C. L., Klein, P., Sun, T. J., Kimmel, A. R., & Devreotes, P. N. (1988). Structure and expression of the cAMP cell-surface receptor. Developmental Genetics, 9(4-5), 227-235.

Structure and expression of the cAMP cell-surface receptor. / Saxe, C. L.; Klein, P.; Sun, T. J.; Kimmel, A. R.; Devreotes, Peter N.

In: Developmental Genetics, Vol. 9, No. 4-5, 1988, p. 227-235.

Research output: Contribution to journalArticle

Saxe, CL, Klein, P, Sun, TJ, Kimmel, AR & Devreotes, PN 1988, 'Structure and expression of the cAMP cell-surface receptor', Developmental Genetics, vol. 9, no. 4-5, pp. 227-235.
Saxe, C. L. ; Klein, P. ; Sun, T. J. ; Kimmel, A. R. ; Devreotes, Peter N. / Structure and expression of the cAMP cell-surface receptor. In: Developmental Genetics. 1988 ; Vol. 9, No. 4-5. pp. 227-235.
@article{ec4d9c3a2a9d496cb35ea032354cf25b,
title = "Structure and expression of the cAMP cell-surface receptor",
abstract = "Using antibodies specific for the 3',5'-cyclic AMP (cAMP) cell surface receptor of Dictyostelium discoideum, we have screened λgtll expression libraries and isolated a series of cDNAs derived from cAMP receptor mRNA during early development. The identity of the cDNA clones was verified by multiple criteria: 1) β-galactosidase fusion proteins synthesized by isolated cDNA clones stain intensely with cAMP receptor directed antiserum, 2) these fusion proteins affinity purify antibodies specific for the cAMP receptor, 3) the cDNA probes hybridize to a 2 kb mRNA whose change in relative level of abundance during development parallels that of receptor mRNA as assayed by in vitro translation, 4) the 2 kb mRNA size equals that of receptor mRNA as determined by in vitro translation of size fractionated poly (A)+ RNA, and 5) RNA transcribed in vitro from cDNAs containing the entire protein-coding region produces a polypeptide by in vitro translation with an apparent molecular weight in close agreement with that of nascent cAMP receptor protein produced by in vitro translation of cellular RNA. The DNA sequence predicts an open reading frame of 392 amino acids. The deduced amino acid sequence contains seven domains enriched in hydrophobic residues. A model is proposed in which the cAMP cell-surface receptor traverses the lipid bilayer seven times in a pattern similar to that of other receptors, such as rhodopsin, which interact with G-proteins. The structural similarities suggest a gene family of related surface receptors from such evolutionarily diverse species as Dictyostelium, yeast, and mammals.",
author = "Saxe, {C. L.} and P. Klein and Sun, {T. J.} and Kimmel, {A. R.} and Devreotes, {Peter N}",
year = "1988",
language = "English (US)",
volume = "9",
pages = "227--235",
journal = "Genesis",
issn = "1526-954X",
publisher = "Wiley-Liss Inc.",
number = "4-5",

}

TY - JOUR

T1 - Structure and expression of the cAMP cell-surface receptor

AU - Saxe, C. L.

AU - Klein, P.

AU - Sun, T. J.

AU - Kimmel, A. R.

AU - Devreotes, Peter N

PY - 1988

Y1 - 1988

N2 - Using antibodies specific for the 3',5'-cyclic AMP (cAMP) cell surface receptor of Dictyostelium discoideum, we have screened λgtll expression libraries and isolated a series of cDNAs derived from cAMP receptor mRNA during early development. The identity of the cDNA clones was verified by multiple criteria: 1) β-galactosidase fusion proteins synthesized by isolated cDNA clones stain intensely with cAMP receptor directed antiserum, 2) these fusion proteins affinity purify antibodies specific for the cAMP receptor, 3) the cDNA probes hybridize to a 2 kb mRNA whose change in relative level of abundance during development parallels that of receptor mRNA as assayed by in vitro translation, 4) the 2 kb mRNA size equals that of receptor mRNA as determined by in vitro translation of size fractionated poly (A)+ RNA, and 5) RNA transcribed in vitro from cDNAs containing the entire protein-coding region produces a polypeptide by in vitro translation with an apparent molecular weight in close agreement with that of nascent cAMP receptor protein produced by in vitro translation of cellular RNA. The DNA sequence predicts an open reading frame of 392 amino acids. The deduced amino acid sequence contains seven domains enriched in hydrophobic residues. A model is proposed in which the cAMP cell-surface receptor traverses the lipid bilayer seven times in a pattern similar to that of other receptors, such as rhodopsin, which interact with G-proteins. The structural similarities suggest a gene family of related surface receptors from such evolutionarily diverse species as Dictyostelium, yeast, and mammals.

AB - Using antibodies specific for the 3',5'-cyclic AMP (cAMP) cell surface receptor of Dictyostelium discoideum, we have screened λgtll expression libraries and isolated a series of cDNAs derived from cAMP receptor mRNA during early development. The identity of the cDNA clones was verified by multiple criteria: 1) β-galactosidase fusion proteins synthesized by isolated cDNA clones stain intensely with cAMP receptor directed antiserum, 2) these fusion proteins affinity purify antibodies specific for the cAMP receptor, 3) the cDNA probes hybridize to a 2 kb mRNA whose change in relative level of abundance during development parallels that of receptor mRNA as assayed by in vitro translation, 4) the 2 kb mRNA size equals that of receptor mRNA as determined by in vitro translation of size fractionated poly (A)+ RNA, and 5) RNA transcribed in vitro from cDNAs containing the entire protein-coding region produces a polypeptide by in vitro translation with an apparent molecular weight in close agreement with that of nascent cAMP receptor protein produced by in vitro translation of cellular RNA. The DNA sequence predicts an open reading frame of 392 amino acids. The deduced amino acid sequence contains seven domains enriched in hydrophobic residues. A model is proposed in which the cAMP cell-surface receptor traverses the lipid bilayer seven times in a pattern similar to that of other receptors, such as rhodopsin, which interact with G-proteins. The structural similarities suggest a gene family of related surface receptors from such evolutionarily diverse species as Dictyostelium, yeast, and mammals.

UR - http://www.scopus.com/inward/record.url?scp=0024210672&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024210672&partnerID=8YFLogxK

M3 - Article

C2 - 3243022

AN - SCOPUS:0024210672

VL - 9

SP - 227

EP - 235

JO - Genesis

JF - Genesis

SN - 1526-954X

IS - 4-5

ER -