Rab-GTPase-dependent endocytic recycling of Kv1.5 in atrial myocytes.
The number of ion channels expressed on the cell surface shapes the complex electrical response of excitable cells. Maintaining a balance between anterograde and retrograde trafficking of channel proteins is vital in regulating steady-state cell surface expression. Kv1.5 is an important voltage-gated K(+) channel in the cardiovascular system underlying the ultra-rapid rectifying potassium current (Ik(ur)), a major repolarizing current in atrial myocytes, and regulating the resting membrane potential and excitability of smooth muscle cells. Defects in the expression of Kv1.5 are associated with pathological states such as chronic atrial fibrillation and hypoxic pulmonary hypertension. There is, thus, substantial interest in understanding the mechanisms regulating cell surface channel levels. Here, we investigated the internalization and recycling of Kv1.5 in the HL-1 immortalized mouse atrial myocytes. Kinetic studies indicate that Kv1.5 is rapidly internalized to a perinuclear region where it co-localizes with the early endosomal marker, EEA1. Importantly, we identified that a population of Kv1.5, originating on the cell surface, internalized and recycled back to the plasma membrane. Notably, Kv1.5 recycling processes are driven by specific Rab-dependent endosomal compartments. Thus, co-expression of GDP-locked Rab4S22N and Rab11S25N dominant-negative mutants decreased the steady-state Kv1.5 surface levels, whereas GTPase-deficient Rab4Q67L and Rab11Q70L mutants increased steady-state Kv1.5 surface levels. These data reveal an unexpected dynamic trafficking of Kv1.5 at the myocyte plasma membrane and demonstrate a role for recycling in the maintenance of steady-state ion channel surface levels.
Department of Pharmacology, University of Michigan, Ann Arbor, Michigan 48109, USA.
Dyke P
DP
McEwen
Sarah M
SM
Schumacher
Qiuju
Q
Li
Mark D
MD
Benson
Jorge A
JA
Iñiguez-Lluhí
Kristin M
KM
Van Genderen
Jeffrey R
JR
Martens
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis; D005799:Genes, Dominant; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006325:Heart Atria / Q000166:cytology; D047468:Immunoprecipitation; D051666:Kv1.5 Potassium Channel / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008954:Models, Biological; D032342:Muscle Cells / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism; D013997:Time Factors; D020691:rab GTP-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D000099193:rab11 GTP-Binding Proteins
D049452:Green Fluorescent Proteins; D051666:Kv1.5 Potassium Channel; D015221:Potassium Channels; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins