Tyrosine phosphorylation of 3BP2 regulates B cell receptor-mediated activation of NFAT.
Adaptor protein c-Abl SH3 domain-binding protein-2 (3BP2, also referred to SH3BP2) regulates immune receptor-mediated signal transduction. In this report we focused on the molecular mechanism of 3BP2 function in B cell receptor (BCR) signaling. Engagement of BCR induces tyrosine phosphorylation of 3BP2. Genetic analysis demonstrated that Syk is critical for BCR-mediated tyrosine phosphorylation of 3BP2. Mutational analysis of 3BP2 revealed that both Tyr(183) and Src homology 2 (SH2) domain are necessary for 3BP2-mediated BCR-induced activation of nuclear factor of activated T cells (NFAT). Point mutation of Tyr(183) or Arg(486) in the SH2 domain of 3BP2 diminished BCR-mediated tyrosine phosphorylation of 3BP2. Endogenous 3BP2 forms a complex with tyrosine-phosphorylated cellular signaling molecules. Peptide binding experiments demonstrated that only phosphorylated Tyr(183) in 3BP2 could form a complex with the SH2 domain(s) of phospholipase Cgamma2 and Vav1 from B cell lysates. These interactions were represented by using bacterial glutathione S-transferase-phospholipase Cgamma2 or -Vav1 SH2 domain. Furthermore, pulldown and Far Western experiments showed that the 3BP2-SH2 domain directly binds to B cell linker protein (BLNK) after BCR stimulation. These results demonstrated that 3BP2 induces the protein complex with cellular signaling molecules through phosphorylation of Tyr(183) and SH2 domain leading to the activation of NFAT in B cells.
Shukla
Upasana
U
Division of Microbiology, Department of Pathological Sciences, Faculty of Medical Sciences, University of Fukui, Fukui 910-1193, Japan.
Hatani
Tomoko
T
Nakashima
Kenji
K
Ogi
Kazuhiro
K
Sada
Kiyonao
K
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D002645:Chickens; D004252:DNA Mutational Analysis; D017930:Genes, Reporter; D006801:Humans; D050778:NFATC Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D051562:Proto-Oncogene Proteins c-bcr / Q000378:metabolism; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D050778:NFATC Transcription Factors; D010455:Peptides; D011947:Receptors, Antigen, B-Cell; C109073:SH3BP2 protein, human; D014443:Tyrosine; D051562:Proto-Oncogene Proteins c-bcr