Proteomics

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Identification of immunoglobulin switch (S) region binding proteins by insertional ChIP (iChIP)

ABSTRACT: We used insertional ChIP (iChIP) based approach to identify proteins that bind to mouse immunoglobulin switch (S) region. To perform iChIP, we inserted an 8X-repeat of the LexA-binding element (LexA-BE) downstream of the Sα region in CH12F3-2A cells, a mouse B-cell line. The DNA-binding domain and dimerization domain of the LexA protein fused with a FLAG tag and a nuclear localization signal (NLS) was expressed in WT (FNLDD-alone) or Sα-engineered (FNLDD-Sα-LexA) CH12F3-2A cells. The resultant cells were subjected to stable isotope labeling with amino acids in cell culture (SILAC), stimulated with CIT (CD40L, IL4, and TGFβ), immunoprecipitated with an anti-FLAG antibody, and subjected to LC-MS/MS analysis. The identities of the deposited data are as follows: F: WT (FNLDD-alone); D: Sα-engineered (FNLDD-Sα-LexA).

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): B Cell

SUBMITTER: Fuminori Sugihara  

LAB HEAD: Fuminori Sugihara

PROVIDER: PXD019042 | Pride | 2020-06-15

REPOSITORIES: Pride

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150501_TFujita_F-D_1_1_4u.raw Raw
150501_TFujita_F-D_2_1_4u.raw Raw
150501_TFujita_F-D_3_1_4u.raw Raw
150501_TFujita_F-D_4_1_4u.raw Raw
150501_TFujita_F-D_5_1_4u.raw Raw
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Publications

SAMHD1-mediated dNTP degradation is required for efficient DNA repair during antibody class switch recombination.

Husain Afzal A   Xu Jianliang J   Fujii Hodaka H   Nakata Mikiyo M   Kobayashi Maki M   Wang Ji-Yang JY   Rehwinkel Jan J   Honjo Tasuku T   Begum Nasim A NA  

The EMBO journal 20200608 15

Sterile alpha motif and histidine-aspartic acid domain-containing protein 1 (SAMHD1), a dNTP triphosphohydrolase, regulates the levels of cellular dNTPs through their hydrolysis. SAMHD1 protects cells from invading viruses that depend on dNTPs to replicate and is frequently mutated in cancers and Aicardi-Goutières syndrome, a hereditary autoimmune encephalopathy. We discovered that SAMHD1 localizes at the immunoglobulin (Ig) switch region, and serves as a novel DNA repair regulator of Ig class s  ...[more]

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