Mpl traffics to the cell surface through conventional and unconventional routes.

Traffic 2014 Sep 18;15(9):961-82. Epub 2014 Jul 18.

Department of Pathology & Cancer Center, University of New Mexico Health Sciences Center, Albuquerque, NM, 87131, USA; INSERM UMR892/CNRS UMR6299, Centre de Recherche en Cancérologie Nantes-Angers (CRCNA), Institut de Recherche Thérapeutique-Université de Nantes, Nantes, 44000, France.

Myeloproliferative neoplasms (MPNs) are often characterized by JAK2 or calreticulin (CALR) mutations, indicating aberrant trafficking in pathogenesis. This study focuses on Mpl trafficking and Jak2 association using two model systems: human erythroleukemia cells (HEL; JAK2V617F) and K562 myeloid leukemia cells (JAK2WT). Consistent with a putative chaperone role for Jak2, Mpl and Jak2 associate on both intracellular and plasma membranes (shown by proximity ligation assay) and siRNA-mediated knockdown of Jak2 led to Mpl trapping in the endoplasmic reticulum (ER). Even in Jak2 sufficient cells, Mpl accumulates in punctate structures that partially colocalize with ER-tracker, the ER exit site marker (ERES) Sec31a, the autophagy marker LC3 and LAMP1. Mpl was fused to miniSOG, a genetically encoded tag for correlated light and electron microscopy. Results suggest that a fraction of Mpl is taken up into autophagic structures from the ER and routed to autolyososomes. Surface biotinylation shows that both immature and mature Mpl reach the cell surface; in K562 cells Mpl is also released in exosomes. Both forms rapidly internalize upon ligand addition, while recovery is primarily attributed to immature Mpl. Mpl appears to reach the plasma membrane via both conventional ER-Golgi and autolysosome secretory pathways, as well as recycling.

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http://dx.doi.org/10.1111/tra.12185DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4141020PMC
September 2014
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