Low affinity binding sites in an activating CRM mediate negative autoregulation of the Drosophila Hox gene Ultrabithorax


Autoři: Rebecca K. Delker aff001;  Vikram Ranade aff003;  Ryan Loker aff003;  Roumen Voutev aff001;  Richard S. Mann aff001
Působiště autorů: Department of Biochemistry and Molecular Biophysics and Systems Biology, Columbia University, New York, NY, United States of America aff001;  Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, United States of America aff002;  Department of Genetics, Columbia University, New York, NY, United States of America aff003
Vyšlo v časopise: Low affinity binding sites in an activating CRM mediate negative autoregulation of the Drosophila Hox gene Ultrabithorax. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008444
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008444

Souhrn

Specification of cell identity and the proper functioning of a mature cell depend on precise regulation of gene expression. Both binary ON/OFF regulation of transcription, as well as more fine-tuned control of transcription levels in the ON state, are required to define cell types. The Drosophila melanogaster Hox gene, Ultrabithorax (Ubx), exhibits both of these modes of control during development. While ON/OFF regulation is needed to specify the fate of the developing wing (Ubx OFF) and haltere (Ubx ON), the levels of Ubx within the haltere differ between compartments along the proximal-distal axis. Here, we identify and molecularly dissect the novel contribution of a previously identified Ubx cis-regulatory module (CRM), anterobithorax (abx), to a negative auto-regulatory loop that decreases Ubx expression in the proximal compartment of the haltere as compared to the distal compartment. We find that Ubx, in complex with the known Hox cofactors, Homothorax (Hth) and Extradenticle (Exd), acts through low-affinity Ubx-Exd binding sites to reduce the levels of Ubx transcription in the proximal compartment. Importantly, we also reveal that Ubx-Exd-binding site mutations sufficient to result in de-repression of abx activity in a transgenic context are not sufficient to de-repress Ubx expression when mutated at the endogenous locus, suggesting the presence of multiple mechanisms through which Ubx-mediated repression occurs. Our results underscore the complementary nature of CRM analysis through transgenic reporter assays and genome modification of the endogenous locus; but, they also highlight the increasing need to understand gene regulation within the native context to capture the potential input of multiple genomic elements on gene control.

Klíčová slova:

Alleles – Cloning – DAPI staining – DNA transcription – Drosophila melanogaster – Genetic loci – Transcriptional control – Imaginal discs


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Genetika Reprodukční medicína

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PLOS Genetics


2019 Číslo 10

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