Title | Duf89 abets lncRNA control of fission yeast phosphate homeostasis via its antagonism of precocious lncRNA transcription termination. |
Publication Type | Journal Article |
Year of Publication | 2023 |
Authors | Sanchez AM, Garg A, Schwer B, Shuman S |
Journal | RNA |
Volume | 29 |
Issue | 6 |
Pagination | 808-825 |
Date Published | 2023 Jun |
ISSN | 1469-9001 |
Keywords | Homeostasis, Phosphates, RNA Polymerase II, RNA, Long Noncoding, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Transcription Termination, Genetic, Transcription, Genetic |
Abstract | Fission yeast phosphate homeostasis gene pho1 is actively repressed during growth in phosphate-rich medium by transcription in cis of a long noncoding (lnc) RNA from the 5' flanking prt(nc-pho1) gene. Pho1 expression is: (i) derepressed by genetic maneuvers that favor precocious lncRNA 3'-processing and termination, in response to DSR and PAS signals in prt; and (ii) hyperrepressed in genetic backgrounds that dampen 3'-processing/termination efficiency. Governors of 3'-processing/termination include the RNA polymerase CTD code, the CPF (cleavage and polyadenylation factor) complex, termination factors Seb1 and Rhn1, and the inositol pyrophosphate signaling molecule 1,5-IP8 Here, we present genetic and biochemical evidence that fission yeast Duf89, a metal-dependent phosphatase/pyrophosphatase, is an antagonist of precocious 3'-processing/termination. We show that derepression of pho1 in duf89Δ cells correlates with squelching the production of full-length prt lncRNA and is erased or attenuated by: (i) DSR/PAS mutations in prt; (ii) loss-of-function mutations in components of the 3'-processing and termination machinery; (iii) elimination of the CTD Thr4-PO4 mark; (iv) interdicting CTD prolyl isomerization by Pin1; (v) inactivating the Asp1 kinase that synthesizes IP8; and (vi) loss of the putative IP8 sensor Spx1. The findings that duf89Δ is synthetically lethal with pho1-derepressive mutations CTD-S7A and aps1Δ-and that this lethality is rescued by CTD-T4A, CPF/Rhn1/Pin1 mutations, and spx1Δ-implicate Duf89 more broadly as a collaborator in cotranscriptional regulation of essential fission yeast genes. The duf89-D252A mutation, which abolishes Duf89 phosphohydrolase activity, phenocopied duf89 +, signifying that duf89Δ phenotypes are a consequence of Duf89 protein absence, not absence of Duf89 catalysis. |
DOI | 10.1261/rna.079595.123 |
Alternate Journal | RNA |
PubMed ID | 36882296 |
PubMed Central ID | PMC10187668 |
Grant List | P30 CA008748 / CA / NCI NIH HHS / United States R01 GM134021 / GM / NIGMS NIH HHS / United States R35 GM126945 / GM / NIGMS NIH HHS / United States |
Submitted by ljc4002 on August 21, 2025 - 1:48pm