ATP11C is critical for the internalization of phosphatidylserine and differentiation of B lymphocytes

Abstract

Subcompartments of the plasma membrane are believed to be critical for lymphocyte responses, but few genetic tools are available to test their function. Here we describe a previously unknown X-linked B cell–deficiency syndrome in mice caused by mutations in Atp11c, which encodes a member of the P4 ATPase family thought to serve as 'flippases' that concentrate aminophospholipids in the cytoplasmic leaflet of cell membranes. Defective ATP11C resulted in a lower rate of phosphatidylserine translocation in pro-B cells and much lower pre-B cell and B cell numbers despite expression of pre-rearranged immunoglobulin transgenes or enforced expression of the prosurvival protein Bcl-2 to prevent apoptosis and abolished pre-B cell population expansion in response to a transgene encoding interleukin 7. The only other abnormalities we noted were anemia, hyperbilirubinemia and hepatocellular carcinoma. Our results identify an intimate connection between phospholipid transport and B lymphocyte function.

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Figure 1: Identification and initial characterization of strains with fewer B lymphocytes after N-ethyl-N-nitrosourea–induced mutagenesis.
Figure 2: Identification of a splice-site mutation in Atp11c by next-generation DNA sequencing.
Figure 3: The Atp11camb mutation results in smaller B cell subsets except marginal zone B cells.
Figure 4: Effects of transgenes encoding Bcl-2, IL-7 or the BCR on the development of ATP11C-mutant B cells.
Figure 5: Less transition of pro-B cells from immunoglobulin-negative to immunoglobulin-positive cells and absence of a response to the H2Ea-Il7 transgene despite higher expression of IL-7Rα.
Figure 6: The Atp11camb mutation results in less translocation of PS into pro-B cells.

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Acknowledgements

We thank the staff of the Australian National University Bioscience Research Services for animal husbandry; the Australian Phenomics Facility genotyping and mapping team for genetic analysis; D. Howard for help with the bone marrow chimeras; A. Bröer for help with the flippase assay; staff of the John Curtin School of Medical Research microscopy and flow cytometry resource facility for help with flow cytometry; the Biomolecular Resource Facility of the Australian Cancer Research Foundation and the Australian Genome Research Facility for DNA sequencing; and K. Peng (Biomolecular Resource Facility of the Australian Cancer Research Foundation), John Curtin School of Medical Research and the Australian Genome Research Facility Next Generation Sequencing and Bioinformatics teams, Brisbane, for preparing and running samples in parallel for next-generation sequencing. Supported by the National Institutes of Health (C.C.G.), Wellcome Trust (C.C.G.), the Australian Research Council (C.C.G.), the National Health and Medical Research Council (C.C.G.), the Ramaciotti Foundation (C.C.G. and A.E.), the Ministry of National Education, Republic of Turkey (M.Y.) and Deutsche Forschungsgemeinschaft (EN 790/1-1 to A.E.).

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M.Y. did and analyzed most of the experiments; C.E.T., S.F., D.L. E.M.K., J.K. and A.E. contributed to experiments; C.M.R. and A.E. identified the ATP11C-mutant strains; B.W., D.T.A., Y.Z. and A.E. mapped and identified the mutation in the ambrosius strain; N.C.T. and G.C.F. analyzed liver histology and clinical chemistry; S.B. helped with the flippase assay; C.C.G. and A.E. conceived of the research and directed the study; and M.Y., C.C.G. and A.E. prepared the figures and wrote the paper in consultation with all authors.

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Correspondence to Christopher C Goodnow or Anselm Enders.

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Yabas, M., Teh, C., Frankenreiter, S. et al. ATP11C is critical for the internalization of phosphatidylserine and differentiation of B lymphocytes. Nat Immunol 12, 441–449 (2011). https://doi.org/10.1038/ni.2011

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