Oncomelania hupensis hupensis isolate:GD
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ABSTRACT:
PROVIDER: PRJNA552664 | ENA |
REPOSITORIES: ENA
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Project description:The retention of galactose-deficient IgA1 (Gd-IgA1) in the glomerular mesangium is a hallmark of IgA nephropathy (IgAN). The immune complex formed by Gd-IgA1 binds to transferrin receptor 1 (TfR1) and deposits on mesangial cell, inducing cell proliferation, inflammatory response, matrix secretion, etc., causes organic damage and dysfunction of the kidney. Even though, the mechanism of how Gd-IgA1 recognize and bind to TfR1 is unclear. In this experiment, we ami to determine the specific interaction region responsible for Gd-IgA1 and TfR1. We prepared a complex of Gd-IgA1 and TfR1 by incubating the two proteins under acidic conditions and stabilizing it using the chemical cross-linking agent glutaraldehyde. And then, the gel bands corresponding to the Gd-IgA1-TfR1 complex was exexcised from the SDS-PAGE gel and analyzed by high-resolution mass spectrometry. This analysis identified fragments containing the hinge peptide sequence, with the R276 residue interacting with the E350 and D356 residues in the apical domain of TfR1. Our results reveal that the hinge peptide of Gd-IgA1 interacts specifically with the apical domain of TfR1.
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Project description:We performed chromatin immunoprecipitation followed by high-throughput DNA sequencing (ChIP-seq) for ICN1 in thymic gd T cells from neonates of C57BL/6 mice. Sequence reads for ICN1 and Input were aligned with the mouse reference genome (mm10) by bowtie program (v1.0.0), and peak detection and identification of ICN1 binding sites were obtained with MACS program (v1.4.2). ChIP-seq data revealed that 12%, 6% and 47% of ICN1 binding sites were within -10kb upstream of transcription start site, +10kb downstream of 3'end and the bodies of genes, respectively. The other of ICN1 binding sites was intergenic. We also found that ICN1 directly bound -5.4kb upstream of transcript start site of IL-7 receptor alpha (IL-7Ra) locus, which were estimated as a new promoter region of IL-7Ra. Some of ICN1 binding sites (e.g. TCF1 promoter) were consistent with previous reports, but others (e.g. Nr4a1 promoter) were novel. These suggests that ICN1 was associated with a lot of transcriptional regulation in gd T cells. Examination of ICN1 binding sites in gd T cells
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