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JCS ePress online publication date 15 Mar 2005
doi: 10.1242/jcs.01733

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Research Article

Connexin interaction patterns in keratinocytes revealed morphologically and by FRET analysis


Wei-Li Di, Yan Gu, John E.A. Common, Trond Aasen, Edel A. O'Toole, David P. Kelsell, and Daniel Zicha*
* Author for correspondence (e-mail: daniel.zicha{at}cancer.org.uk)

Multiple connexins, the major proteins of gap junctions, have overlapping expression in the human epidermis and are postulated to have a key role in keratinocyte differentiation and homeostasis. The functional importance of connexins in the epidermis is emphasised by the association of mutations in four human connexins with various hyperproliferative skin disorders. As immunohistochemistry demonstrated overlapping expression of specific connexins in keratinocytes, we performed colocalisation analyses and applied a modified FRET methodology to assess possible heteromeric interactions between different combinations of four wild-type (wt) and mutant connexins. The data generated indicate that there is evidence for multiple connexin interactions at the plasma membrane between (wt)Cx26, (wt)Cx30 and (wt)Cx31 in keratinocytes and thus, the potential for the formation of a large number of different channel types each with different channel properties. In addition, we demonstrate that the inherent in vitro trafficking defect of the skin disease mutations (D50N)Cx26 and (G11R)Cx30 can be overcome partially by the coexpression of different wild-type connexins but this rescue does not result in large gap junction aggregates at the plasma membrane. These data indicate that skin disease associated Cx26 or Cx30 mutations are likely to disrupt a number of different channel types important in distinct aspects of keratinocyte biology.
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