Identification of Human Pathways Acting on Nuclear Non-Coding RNAs Using the Mirror Forward Genetic Approach.

Autor:	Che R; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA.; Clemson University Center for Human Genetics, Greenwood, SC 29646, USA., Panah M; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA.; Clemson University Center for Human Genetics, Greenwood, SC 29646, USA., Mirani B; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA.; Clemson University Center for Human Genetics, Greenwood, SC 29646, USA., Knowles K; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA.; Clemson University Center for Human Genetics, Greenwood, SC 29646, USA., Ostapovich A; Dept. of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06536, USA., Majumdar D; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA.; Clemson University Center for Human Genetics, Greenwood, SC 29646, USA., Chen X; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA., DeSimone J; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA., White W; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA., Noonan M; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA., Luo H; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA., Alexandrov A; Dept. of Genetics and Biochemistry, Clemson University, Clemson, SC 29631, USA.; Clemson University Center for Human Genetics, Greenwood, SC 29646, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Sep 27. Date of Electronic Publication: 2024 Sep 27.
DOI:	10.1101/2024.09.26.615073
Abstrakt:	Despite critical roles in diseases, human pathways acting on strictly nuclear non-coding RNAs have been refractory to forward genetics. To enable their forward genetic discovery, we developed a single-cell approach that "Mirrors" activities of nuclear pathways with cytoplasmic fluorescence. Application of Mirror to two nuclear pathways targeting MALAT1's 3' end, the pathway of its maturation and the other, the degradation pathway blocked by the triple-helical Element for Nuclear Expression (ENE), identified nearly all components of three complexes: Ribonuclease P and the RNA Exosome, including nuclear DIS3, EXOSC10, and C1D, as well as the Nuclear Exosome Targeting (NEXT) complex. Additionally, Mirror identified DEAD-box helicase DDX59 associated with the genetic disorder Oral-Facial-Digital syndrome (OFD), yet lacking known substrates or roles in nuclear RNA degradation. Knockout of DDX59 exhibits stabilization of the full-length MALAT1 with a stability-compromised ENE and increases levels of such long non-coding RNAs as NEAT1_1 and NIPBL-DT, as well as 3'-extended forms of small nuclear RNAs. It also exhibits extensive retention of minor introns, including in OFD-associated genes, suggesting a mechanism for DDX59 association with OFD. Mirror efficiently identifies pathways acting on strictly nuclear non-coding RNAs, including essential and indirectly-acting components, and, as a result, uncovers unexpected links to human disease. Competing Interests: COMPETING INTERESTS The authors declare no competing interests.
Databáze:	MEDLINE
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