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. 2014 May 1;28(9):959–970. doi: 10.1101/gad.236729.113

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© 2014 Chen and Gartenberg; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 3. — tRNA synthesis during the cell cycle. (A) Rpc25 occupancy at tDNAs. ChIP-qPCR of Rpc25-TAP was performed with strain MC195 arrested in M phase or G1. Rpc25 binding at tDNAs was enriched significantly during M-phase arrest (P-values are in Supplemental Table S4). (B) Pre-tRNA^Ser(CGA) of S. cerevisiae and S. bayanus. Red circles represent the conserved intronic nucleotides, and green circles represent intronic nucleotide polymorphisms. Primers for RT (primer 1) and qPCR (primers 1 and 2) are shown. (C) Nascent tRNA levels. RT-qPCR was performed with strain MC204 [Δtt(agu)c∷Sb-tS(CGA)C]. tRNA levels were significantly higher in M phase (P-values are in Supplemental Table S4). (D) Northern blot analysis of tRNA precursors and products. Radiolabeled oligonucleotide probes were hybridized to unspliced and spliced tRNA^Ser(CGA), albeit with different affinities, in extracts of strain MC172. Unprocessed, 5′ processed, end processed, and mature species are labeled graphically. The ratio of end processed/spliced species is indicated below each lane.