HAP1 helps to regulate actin-based transport of insulin-containing granules in pancreatic β cells.
Huntingtin-associated protein 1 (HAP1) is enriched in neurons and binds to polyglutamine-expanded huntingtin. It consists of two alternatively spliced isoforms, HAP1A and HAP1B, which differ only in their short C-terminal sequences. Both HAP1A and HAP1B have been also detected in pancreatic β cells, where the loss of HAP1 impairs glucose-stimulated insulin secretion. Here, we use time-lapse laser scanning confocal microscopy to provide direct evidence that HAP1A, but not HAP1B, co-localizes and co-migrates with insulin-containing vesicles and actin-based myosin Va motor protein in the INS-1 pancreatic β cell line. Knocking down HAP1 expression using small interfering RNA significantly inhibited actin-based transport of insulin vesicles following glucose stimulation. Co-immunoprecipitation experiments demonstrated interaction between HAP1A, myosin Va, and phogrin, a transmembrane protein in insulin-containing vesicles. Stimulating INS-1 cells with glucose increased the association of HAP1A with myosin Va, while silencing HAP1 expression reduced the association of myosin Va with phogrin after glucose stimulation, without affecting levels of myosin Va or actin. Our results provide real-time evidence in living cells that HAP1 may help regulate transport of insulin-containing secretory granules along cortical actin filaments. This also raises the possibility that HAP1 may play an important role in actin-based secretory vesicle trafficking in neurons.
Division of Histology and Embryology, Department of Anatomy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People's Republic of China, wanliheng@vip.163.com.
Zhiyong
Z
Wang
Ting
T
Peng
Hongnian
H
Wu
Jun
J
He
He
H
Li
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D047468:Immunoprecipitation; D007328:Insulin / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000378:metabolism; D024701:Myosin Type V / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D051381:Rats; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8 / Q000378:metabolism; D022142:Secretory Vesicles / Q000378:metabolism*
C490757:Hap1 protein, rat; D007328:Insulin; C513081:Myo5a protein, rat; D009419:Nerve Tissue Proteins; C105576:Ptprn2 protein, rat; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D024701:Myosin Type V; D018995:Myosin Heavy Chains