ATP depletion alters myosin I beta cellular location in LLC-PK1 cells.
The brush border (BB) of the proximal tubule cell (PTC) requires dynamic membrane events for function. The actin cytoskeleton is necessary for structure and function in this region. ATP depletion disrupts both structure and function. In this report, myosin 1 beta location in LLC-PK1 cells was followed during ATP depletion and repletion using immunofluorescence and Western blot techniques. Myosin I beta colocalized with F-actin in the microvilli and cell periphery, but no colocalization was observed with stress fibers. ATP depletion increased the apical F-actin, and myosin I beta was colocalized there. In addition, after ATP depletion, myosin I beta was extracted less by Triton X-100. These changes were reversed after ATP repletion. Finally, immunofluorescence of kidney sections shows myosin I beta in the BB. These results place this motor in a dynamic region of the PTC where its actin and membrane binding domains can contribute to PTC function.
Wagner
M C
MC
Department of Medicine, Indiana University Medical Center, Indianapolis 46202, USA.
Molitoris
B A
BA
D000199:Actins / Q000502:physiology; D000255:Adenosine Triphosphate / Q000172:deficiency* / Q000378:metabolism; D000818:Animals; D003432:Cross-Linking Reagents; D007668:Kidney / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000201:enzymology; D018374:LLC-PK1 Cells / Q000378:metabolism*; D008871:Microvilli / Q000201:enzymology; D009218:Myosins / Q000378:metabolism* / Q000502:physiology; D017830:Octoxynol; D051381:Rats; D012016:Reference Values; D012995:Solubility; D013552:Swine; D014018:Tissue Distribution
D000199:Actins; D003432:Cross-Linking Reagents; D000255:Adenosine Triphosphate; D017830:Octoxynol; D009218:Myosins