FASTKD1 and FASTKD4 have opposite effects on expression of specific mitochondrial RNAs, depending upon their endonuclease-like RAP domain.

Autor:	Boehm E; Cell Biology Department, University of Geneva, 1211 Geneva 4, Switzerland., Zaganelli S; Cell Biology Department, University of Geneva, 1211 Geneva 4, Switzerland., Maundrell K; Cell Biology Department, University of Geneva, 1211 Geneva 4, Switzerland., Jourdain AA; Cell Biology Department, University of Geneva, 1211 Geneva 4, Switzerland., Thore S; INSERM U-1212, CNRS UMR 5320, Université de Bordeaux, ARNA Laboratory, Bordeaux 33000, France., Martinou JC; Cell Biology Department, University of Geneva, 1211 Geneva 4, Switzerland.
Jazyk:	angličtina
Zdroj:	Nucleic acids research [Nucleic Acids Res] 2017 Jun 02; Vol. 45 (10), pp. 6135-6146.
DOI:	10.1093/nar/gkx164
Abstrakt:	FASTK family proteins have been identified as regulators of mitochondrial RNA homeostasis linked to mitochondrial diseases, but much remains unknown about these proteins. We show that CRISPR-mediated disruption of FASTKD1 increases ND3 mRNA level, while disruption of FASTKD4 reduces the level of ND3 and of other mature mRNAs including ND5 and CYB, and causes accumulation of ND5-CYB precursor RNA. Disrupting both FASTKD1 and FASTKD4 in the same cell results in decreased ND3 mRNA similar to the effect of depleting FASTKD4 alone, indicating that FASTKD4 loss is epistatic. Interestingly, very low levels of FASTKD4 are sufficient to prevent ND3 loss and ND5-CYB precursor accumulation, suggesting that FASTKD4 may act catalytically. Furthermore, structural modeling predicts that each RAP domain of FASTK proteins contains a nuclease fold with a conserved aspartate residue at the putative active site. Accordingly, mutation of this residue in FASTKD4 abolishes its function. Experiments with FASTK chimeras indicate that the RAP domain is essential for the function of the FASTK proteins, while the region upstream determines RNA targeting and protein localization. In conclusion, this paper identifies new aspects of FASTK protein biology and suggests that the RAP domain function depends on an intrinsic nucleolytic activity. (© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.)
Databáze:	MEDLINE
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