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2014 ; 159
(1
): 148-162
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Transcriptome-wide mapping reveals widespread dynamic-regulated pseudouridylation
of ncRNA and mRNA
#MMPMID25219674
Schwartz S
; Bernstein DA
; Mumbach MR
; Jovanovic M
; Herbst RH
; León-Ricardo BX
; Engreitz JM
; Guttman M
; Satija R
; Lander ES
; Fink G
; Regev A
Cell
2014[Sep]; 159
(1
): 148-162
PMID25219674
show ga
Pseudouridine is the most abundant RNA modification, yet except for a few
well-studied cases, little is known about the modified positions and their
function(s). Here, we develop ?-seq for transcriptome-wide quantitative mapping
of pseudouridine. We validate ?-seq with spike-ins and de novo identification of
previously reported positions and discover hundreds of unique sites in human and
yeast mRNAs and snoRNAs. Perturbing pseudouridine synthases (PUS) uncovers which
pseudouridine synthase modifies each site and their target sequence features.
mRNA pseudouridinylation depends on both site-specific and snoRNA-guided
pseudouridine synthases. Upon heat shock in yeast, Pus7p-mediated
pseudouridylation is induced at >200 sites, and PUS7 deletion decreases the
levels of otherwise pseudouridylated mRNA, suggesting a role in enhancing
transcript stability. rRNA pseudouridine stoichiometries are conserved but
reduced in cells from dyskeratosis congenita patients, where the PUS DKC1 is
mutated. Our work identifies an enhanced, transcriptome-wide scope for
pseudouridine and methods to dissect its underlying mechanisms and function.