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Three proteins walk into a "var"…

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Antigenic variation is used by the most virulent malaria parasite, Plasmodium falciparum, to evade the human immune system. The most important variant surface antigen is encoded by a family of ~60 var genes that are expressed in a mutually exclusive fashion to ensure a sustained infection. In this epigenetically-regulated process, all var genes are compacted into silent chromatin clusters at the nuclear periphery. Singular var gene activation is concomitant with chromatin decompaction and separation into an ‘activation center’. While the basic epigenetic signatures of silent versus active var genes are known, much remains to be discovered about how they are regulated and organized within the nucleus. To identify new proteins involved in this process, we recently purified the var gene promoter using immunoprecipitation with an enzymatically inactive Cas9 coupled with mass spectrometry. We identified a protein complex including an ApiAP2 transcription factor, a putative microorchidia family protein (MORC), and acetyl CoA synthetase (ACAS). Chromatin immunoprecipitation and sequencing confirmed that these proteins bind together and, more specifically, at the promoters of var genes found in subtelomeric regions. Inducible knockdown followed by differential expression analysis showed that the members of this complex are involved in controlling mutually exclusive expression of the var gene family. Ongoing work seeks to understand the role of these proteins in var gene regulation and organization and investigates a potential link to parasite metabolism.

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