RNAi-mediated depletion of the NSL complex subunits leads to abnormal chromosome segregation and defective centrosome duplication in Drosophila mitosis


Autoři: Gera A. Pavlova aff001;  Julia V. Popova aff001;  Evgeniya N. Andreyeva aff001;  Lyubov A. Yarinich aff001;  Mikhail O. Lebedev aff001;  Alyona V. Razuvaeva aff001;  Tatiana D. Dubatolova aff001;  Anastasiya L. Oshchepkova aff001;  Claudia Pellacani aff005;  Maria Patrizia Somma aff005;  Alexey V. Pindyurin aff001;  Maurizio Gatti aff005
Působiště autorů: Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, Russia aff001;  Institute of Cytology and Genetics, Siberian Branch of RAS, Novosibirsk, Russia aff002;  Novosibirsk State University, Novosibirsk, Russia aff003;  Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of RAS, Novosibirsk, Russia aff004;  IBPM CNR c/o Department of Biology and Biotechnology, Sapienza University of Rome, Rome, Italy aff005
Vyšlo v časopise: RNAi-mediated depletion of the NSL complex subunits leads to abnormal chromosome segregation and defective centrosome duplication in Drosophila mitosis. PLoS Genet 15(9): e32767. doi:10.1371/journal.pgen.1008371
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008371

Souhrn

The Drosophila Nonspecific Lethal (NSL) complex is a major transcriptional regulator of housekeeping genes. It contains at least seven subunits that are conserved in the human KANSL complex: Nsl1/Wah (KANSL1), Dgt1/Nsl2 (KANSL2), Rcd1/Nsl3 (KANSL3), Rcd5 (MCRS1), MBD-R2 (PHF20), Wds (WDR5) and Mof (MOF/KAT8). Previous studies have shown that Dgt1, Rcd1 and Rcd5 are implicated in centrosome maintenance. Here, we analyzed the mitotic phenotypes caused by RNAi-mediated depletion of Rcd1, Rcd5, MBD-R2 or Wds in greater detail. Depletion of any of these proteins in Drosophila S2 cells led to defects in chromosome segregation. Consistent with these findings, Rcd1, Rcd5 and MBD-R2 RNAi cells showed reduced levels of both Cid/CENP-A and the kinetochore component Ndc80. In addition, RNAi against any of the four genes negatively affected centriole duplication. In Wds-depleted cells, the mitotic phenotypes were similar but milder than those observed in Rcd1-, Rcd5- or MBD-R2-deficient cells. RT-qPCR experiments and interrogation of published datasets revealed that transcription of many genes encoding centromere/kinetochore proteins (e.g., cid, Mis12 and Nnf1b), or involved in centriole duplication (e.g., Sas-6, Sas-4 and asl) is substantially reduced in Rcd1, Rcd5 and MBD-R2 RNAi cells, and to a lesser extent in wds RNAi cells. During mitosis, both Rcd1-GFP and Rcd5-GFP accumulate at the centrosomes and the telophase midbody, MBD-R2-GFP is enriched only at the chromosomes, while Wds-GFP accumulates at the centrosomes, the kinetochores, the midbody, and on a specific chromosome region. Collectively, our results suggest that the mitotic phenotypes caused by Rcd1, Rcd5, MBD-R2 or Wds depletion are primarily due to reduced transcription of genes involved in kinetochore assembly and centriole duplication. The differences in the subcellular localizations of the NSL components may reflect direct mitotic functions that are difficult to detect at the phenotypic level, because they are masked by the transcription-dependent deficiency of kinetochore and centriolar proteins.

Klíčová slova:

Biology and life sciences – Genetics – Epigenetics – RNA interference – Gene expression – DNA transcription – Genetic interference – Biochemistry – Nucleic acids – RNA – Cell biology – Cellular structures and organelles – Centrosomes – Centrioles – Cell processes – Cell cycle and cell division – Telophase – Mitosis – Metaphase – Chromosome biology – Organisms – Eukaryota – Animals – Invertebrates – Arthropoda – Insects – Drosophila – Drosophila melanogaster – Research and analysis methods – Animal studies – Experimental organism systems – Model organisms – Animal models


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