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Enhancer bound proteome of intronic Ig Emu enhancer with reverse DNA sequence polarity control bait

ABSTRACT: The specificity of humoral immune responses depends on the functional rearrangement and expression of only one allele of immunoglobulin (Ig) genes. Here, we analyzed the comprehensive proteome of the murine Ig Emu enhancer, which governs the rearrangement and expression of the Ig mu heavy chain allele. By mass spectrometry of proteins bound at wild type versus mutant Emu enhancers, we identified Emu-binding proteins and associated multi-protein complexes. We found that the MSL/MOF complex, a regulator of gene dosage compensation in flies, binds Emu via transcription factor YY1 and facilitates Emu-driven chromatin looping and promoter interaction. Msl2 gene knockout in primary pre-B cells or Mof heterozygosity in mice reduced mu gene expression. In this data set we compare proteins binding to the wild-type Emu versus a DNA bait control for which the entire Emu sequence was switched to its reverse polarity sequence. The latter conserves the DNA GC content but virtually destroys all sequence-specific transcription factor binding sites. Of note, DNA repetitive sequences that can also be bound by DNA interacting proteins are kept functional by this control bait. SILAC quantitative proteomics was employed in a label swap approach incubating wild-type and control DNA with labeled and non-labled protein extracts, respectively.

INSTRUMENT(S): LTQ Orbitrap XL ETD

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): B Cell

DISEASE(S): Lymphoma

SUBMITTER: Gerhard Mittler

LAB HEAD: Gerhard Mittler

PROVIDER: PXD045636 | Pride | 2024-10-25

REPOSITORIES: Pride

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			Action	DRS
	Rep01_EnhReverseSeqFwd01.raw	Raw
	Rep01_EnhReverseSeqFwd02.raw	Raw
	Rep01_EnhReverseSeqFwd03.raw	Raw
	Rep01_EnhReverseSeqFwd04.raw	Raw
	Rep01_EnhReverseSeqFwd05.raw	Raw

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