TRPML cation channels regulate the specialized lysosomal compartment of vertebrate B-lymphocytes.
B-lymphocytes possess a specialized lysosomal compartment, the regulated transformation of which has been implicated in B-cell antigen presentation. Members of the mucolipin (TRPML) family of cation channels have been implicated in regulated vesicular transport in several tissues, but a role for TRPML function in lymphocyte vesicular transport physiology has not been previously described. To address the role of TRPML proteins in lymphocyte vesicular transport, we analyzed the lysosomal compartment in cultured B-lymphocytes engineered to lack TRPML1 or after expression of N- or C-terminal GFP fusion proteins of TRPML1 or TRPML2. Consistent with previous analyses of lymphocytes derived from human patients with mutations in TRPML1, we were not able to detect abnormalities in the lysosomes of TRPML1-deficient DT40 B-lymphocytes. However, while N-terminal GFP fusions of TRPML2 localized to normal appearing lysosomes, C-terminal GFP fusions of either TRPML1 or TRPML2 acted to antagonize endogenous TRPML function, localizing to large vesicular structures, the histological properties of which were indistinguishable from the enlarged lysosomes observed in affected tissues of TRPML1-deficient humans. Endocytosed B-cell receptors were delivered to these enlarged lysosomes, demonstrating that a TRPML-dependent process is required for normal regulation of the specialized lysosome compartment of vertebrate B-lymphocytes.
Song
Yumei
Y
Department of Pediatrics and Immunology, University of Washington and Children's Hospital and Regional Medical Center, Seattle, WA 98195, USA.
Dayalu
Rashmi
R
Matthews
Sharon A
SA
Scharenberg
Andrew M
AM
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D002478:Cells, Cultured; D002645:Chickens; D004705:Endocytosis / Q000502:physiology; D005786:Gene Expression Regulation; D006801:Humans; D008247:Lysosomes / Q000502:physiology* / Q000648:ultrastructure; D051379:Mice; D050053:TRPM Cation Channels / Q000235:genetics / Q000502:physiology*; D050051:Transient Receptor Potential Channels / Q000235:genetics / Q000502:physiology; D022161:Transport Vesicles / Q000502:physiology / Q000648:ultrastructure
C416674:MCOLN1 protein, human; C510641:Mcoln2 protein, human; D050053:TRPM Cation Channels; D050051:Transient Receptor Potential Channels