BCALM (AC099524.1) Is a Human B Lymphocyte-Specific Long Noncoding RNA That Modulates B Cell Receptor-Mediated Calcium Signaling

J Immunol. 2020 Aug 1;205(3):595-607. doi: 10.4049/jimmunol.2000088. Epub 2020 Jun 22.


Of the thousands of long noncoding RNAs (lncRNA) identified in lymphocytes, very few have defined functions. In this study, we report the discovery and functional elucidation of a human B cell-specific lncRNA with high levels of expression in three types of B cell cancer and normal B cells. The AC099524.1 gene is upstream of the gene encoding the B cell-specific phospholipase C γ 2 (PLCG2), a B cell-specific enzyme that stimulates intracellular Ca2+ signaling in response to BCR activation. AC099524.1 (B cell-associated lncRNA modulator of BCR-mediated Ca+ signaling [BCALM]) transcripts are localized in the cytoplasm and, as expected, CRISPR/Cas9 knockout of AC099524.1 did not affect PLCG2 mRNA or protein expression. lncRNA interactome, RNA immunoprecipitation, and coimmunoprecipitation studies identified BCALM-interacting proteins in B cells, including phospholipase D 1 (PLD1), and kinase adaptor proteins AKAP9 (AKAP450) and AKAP13 (AKAP-Lbc). These two AKAP proteins form signaling complexes containing protein kinases A and C, which phosphorylate and activate PLD1 to produce phosphatidic acid (PA). BCR stimulation of BCALM-deficient B cells resulted in decreased PLD1 phosphorylation and increased intracellular Ca+ flux relative to wild-type cells. These results suggest that BCALM promotes negative feedback that downmodulates BCR-mediated Ca+ signaling by promoting phosphorylation of PLD1 by AKAP-associated kinases, enhancing production of PA. PA activates SHP-1, which negatively regulates BCR signaling. We propose the name BCALM for B-Cell Associated LncRNA Modulator of BCR-mediated Ca+ signaling. Our findings suggest a new, to our knowledge, paradigm for lncRNA-mediated modulation of lymphocyte activation and signaling, with implications for B cell immune response and BCR-dependent cancers.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • A Kinase Anchor Proteins / genetics
  • A Kinase Anchor Proteins / immunology
  • Adult
  • Aged
  • Aged, 80 and over
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology*
  • Calcium Signaling / genetics
  • Calcium Signaling / immunology*
  • Cell Line
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / immunology
  • Female
  • Humans
  • Male
  • Middle Aged
  • Minor Histocompatibility Antigens / genetics
  • Minor Histocompatibility Antigens / immunology
  • Phospholipase C gamma / genetics
  • Phospholipase C gamma / immunology
  • Phospholipase D / genetics
  • Phospholipase D / immunology
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6 / genetics
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6 / immunology
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / immunology
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / immunology*
  • Receptors, Antigen, B-Cell / genetics
  • Receptors, Antigen, B-Cell / immunology*


  • A Kinase Anchor Proteins
  • AKAP13 protein, human
  • AKAP9 protein, human
  • Cytoskeletal Proteins
  • Minor Histocompatibility Antigens
  • Proto-Oncogene Proteins
  • RNA, Long Noncoding
  • Receptors, Antigen, B-Cell
  • PTPN6 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • PLCG2 protein, human
  • Phospholipase C gamma
  • Phospholipase D
  • phospholipase D1