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Brem, A.

Publications and source records attributed to Brem, A..

2 recordsLinked to original sources

Sequence-dependent binding modes of INO80 control +1 nucleosome positioning

Cellular self-organisation counteracts entropy at the expenditure of energy. In case of the first level of nuclear DNA organisation, this relates to regular nucleosome arrays and interspaced nucleosome-depleted regions (NDRs), for example at promoters or replication origins1-5. The organisation of nucleosomes as building blocks of chromatin is orchestrated by the collective activities of ATP-dependent chromatin remodellers6-9. Yet, how remodellers achieve positional specificity, in particular regarding the promoter-proximal +1 nucleosomes, remains unclear. Here, we show that the S. cerevisiae chromatin remodeller INO80 unexpectedly distinguishes DNA sequence asymmetry within the +1 nucleosome of the SWH1 gene through distinct inhibited and active nucleosome-binding modes. In structural and biochemical analyses of INO80 on nucleosomes with the endogenous sequence, we identified an inhibited binding mode where the entire INO80 remodelling unit flipped on the +1 nucleosome. INO80 adopted this remodelling-incompetent binding mode when facing the promoter, but a remodelling-competent mode when facing the gene body. This directional read-out of intra-nucleosomal DNA sequence asymmetry, together with extranucleosomal NDR sequence features, prevented nucleosome sliding into the NDR while permitting array formation over the gene. Our work shows how DNA features contribute to ATP-dependent self-organisation of promoter chromatin by INO80.

biochemistry↗

Direct RNA-binding by MYCN mediates feedback from RNA processing to transcription control

The MYCN oncoprotein broadly binds active promoters in a heterodimer with its partner protein MAX. MYCN also interacts with the nuclear exosome, a 3-5 exoribonuclease complex, suggesting a function in RNA metabolism. Here we show that MYCN forms stable high molecular weight complexes with the exosome and multiple RNA-binding proteins. In cells, MYCN binds to thousands of intronic RNAs; recombinant MYCN directly binds RNA via a short, highly conserved sequence termed MYCBoxI. Perturbing exosome function results in global re-localization of MYCN from promoters to intronic RNAs. At promoters, MYCN is then replaced by the MNT(MXD6) repressor protein, which inhibits MYCN-dependent transcription. MYCN promotes the degradation of its bound introns via the nuclear exosome targeting (NEXT) complex. Our data demonstrate that MYCN is an RNA-binding protein that regulates nascent transcript turnover and show that competition between its RNA- and DNA-bound states links the dynamics of the MYCN/MAX/MXD network to mRNA processing.

cancer biology↗