An intact connexin N-terminus is required for function but not gap junction formation.
The cytoplasmic N-termini of connexins have been implicated in protein trafficking, oligomerization and channel gating. To elucidate the role of the N-terminus in connexin37 (CX37), we studied mutant constructs containing partial deletions of its 23 N-terminal amino acids and a construct with a complete N-terminus in which residues 2-8 were replaced with alanines. All mutants containing nine or more N-terminal amino acids form gap junction plaques in transiently transfected HeLa cells, whereas most of the longer deletions do not. Although wild-type CX37 allowed intercellular transfer of microinjected neurobiotin in HeLa cells and formed conducting hemichannels in Xenopus oocytes, none of the mutant constructs tested show evidence of channel function. However, in coexpression experiments, N-terminal mutants that formed gap junction plaques potently inhibit hemichannel conductance of wild-type CX37 suggesting their co-oligomerization. We conclude that as much as half the length of the connexin N-terminus can be deleted without affecting formation of gap junction plaques, but an intact N-terminus is required for hemichannel gating and intercellular communication.
Kyle
John W
JW
Department of Medicine, Section of Cardiology, University of Chicago, Chicago, IL 60637, USA.
Minogue
Peter J
PJ
Thomas
Bettina C
BC
Domowicz
Denise A Lopez
DA
Berthoud
Viviana M
VM
Hanck
Dorothy A
DA
Beyer
Eric C
EC
D000409:Alanine / Q000235:genetics / Q000502:physiology; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000502:physiology; D002450:Cell Communication; D017630:Connexins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D017629:Gap Junctions / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D015640:Ion Channel Gating / Q000235:genetics / Q000502:physiology; D007473:Ion Channels / Q000378:metabolism / Q000502:physiology; D008954:Models, Biological; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000502:physiology; D017384:Sequence Deletion / Q000502:physiology; D014162:Transfection; D000097006:Gap Junction alpha-4 Protein
D000409:Alanine; D017630:Connexins; D007473:Ion Channels; D000097006:Gap Junction alpha-4 Protein