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PMID: 8049527 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Molecular cloning and functional characterization of chick lens fiber connexin 45.6.

Molecular biology of the cell ·Vol. 5 ·No. 3 ·1994-03-00 ·Pages 363-73

Jiang JX, White TW, Goodenough DA, Paul DL

Abstract

The avian lens is an ideal system to study gap junctional intercellular communication in development and homeostasis. The lens is experimentally more accessible in the developing chick embryo than in other organisms, and chick lens cells differentiate well in primary cultures. However, only two members of the connexin gene family have been identified in the avian lens, whereas three are known in the mammalian system. We report here the molecular cloning and characterization of the third lens connexin, chick connexin45.6 (ChCx45.6), a protein with a predicted molecular mass of 45.6 kDa. ChCx45.6 was encoded by a single copy gene and was expressed specifically in the lens. There were two mRNA species of 6.4 kilobase (kb) and 9.4 kb in length. ChCx45.6 was a functional connexin protein, because expression in Xenopus oocyte pairs resulted in the development of high levels of conductance with a characteristic voltage sensitivity. Antisera were raised against ChCx45.6 and chick connexin56 (ChCx56), another avian lens-specific connexin, permitting the examination of the distribution of both proteins. Immunofluorescence localization showed that both ChCx45.6 and ChCx56 were abundant in lens fibers. Treatment of lens membranes with alkaline phosphatase resulted in electrophoretic mobility shifts, demonstrating that both ChCx45.6 and ChCx56 were phosphoproteins in vivo.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Chick Embryo Cloning, Molecular Connexins/genetics,physiology DNA/genetics DNA Primers/genetics Eye Proteins/genetics,physiology Female Gene Expression Lens, Crystalline/physiology Mice Molecular Sequence Data Oocytes/physiology Phosphorylation Sequence Homology, Amino Acid Sheep Species Specificity Tissue Distribution
Chemicals
Connexins DNA Primers Eye Proteins connexin 45 connexin 56 DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Jiang J X
Department of Cell Biology, Neurobiology Harvard Medical School, Boston 02115.
White T W
Goodenough D A
Paul D L
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.5.3.363
Published
1994-03-00
Pages
363-73
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC301043
Subset
IM
Grants
NEI NIH HHS · EY02430 · United States
NEI NIH HHS · F32 EY06448 · United States
NIGMS NIH HHS · GM37751 · United States
Databases
GENBANK
L24799
External Links
PubMed source DOI: 10.1091/mbc.5.3.363 PMC Full Text View journal details
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