Enhanced connexin 43 expression delays intra-mitotic duration and cell cycle traverse independently of gap junction channel function.
Connexins (Cxs) and gap junction (GJ)-mediated communication have been linked with the regulation of cell cycle traverse. However, it is not clear whether Cx expression or GJ channel function are the key mediators in this process or at what stage this regulation may occur. We therefore tested the hypothesis that enhanced Cx expression could alter the rate of cell cycle traverse independently of GJ channel function. Sodium butyrate (NaBu) or anti-arrhythmic peptide (AAP10) were used to enhance Cx expression in HeLa cells stably expressing Cx43 (HeLa-43) and primary cultures of human fibroblasts (HFF) that predominantly express Cx43. To reduce GJ-mediated communication, 18-alpha-glycyrrhetinic acid (GA) was used. In HeLa-43 and HFF cells, NaBu and AAP10 enhanced Cx43 expression and increased channel function, while GA reduced GJ-mediated communication but did not significantly alter Cx43 expression levels. Timelapse microscopy and flow cytometry of HeLa-WT (wild-type, Cx deficient) and HeLa-43 cells dissected cell cycle traverse and enabled measurements of intra-mitotic time and determined levels of G1 arrest. Enhanced Cx43 expression increased mitotic durations corresponding with a G1 delay in cell cycle, which was linked to an increase in expression of the cell cycle inhibitor p21(waf1/cip1) in both HeLa-43 and HFF cells. Reductions in Cx43 channel function did not abrogate these responses, indicating that GJ channel function was not a critical factor in reducing cell proliferation in either cell type. We conclude that enhanced Cx43 expression and not GJ-mediated communication, is involved in regulating cell cycle traverse.
Department of Biological and Biomedical Sciences, School of Life Sciences, Glasgow Caledonian University, 70 Cowcaddens Rd, Glasgow, Scotland G4 0BA, UK.
Scott R
SR
Johnstone
Angela K
AK
Best
Catherine S
CS
Wright
Brant E
BE
Isakson
Rachel J
RJ
Errington
Patricia E
PE
Martin
D015153:Blotting, Western; D002453:Cell Cycle / Q000502:physiology*; D002469:Cell Separation; D018031:Connexin 43 / Q000378:metabolism*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D017629:Gap Junctions / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008853:Microscopy; D008938:Mitosis / Q000502:physiology*; D013995:Time
D018031:Connexin 43