Display Settings:

Items per page

Results: 6

1.
FIGURE 6.

FIGURE 6. From: tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae.

m5C increases specifically on tRNAHis during starvation. tRNAHis (black), tRNAVal(AAC) (dark gray), and tRNAGly(GCC) (light gray) were purified from BY4741 grown in the indicated starvation conditions. Nucleoside modifications were analyzed by HPLC, and m5C values are plotted over time for each tRNA.

Melanie A. Preston, et al. RNA. 2013 February;19(2):243-256.
2.
FIGURE 2.

FIGURE 2. From: tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae.

Temperature-sensitive strains have additional m5C on tRNAHis when grown at nonpermissive temperature. (A) Assessment of viable cell titer following growth at 37°C. (B) m5C levels in tRNAHis and tRNAVal(AAC) during growth at 37°C. tRNAHis and tRNAVal(AAC) were purified from BY4741, thg1-Y146H, and fcp1-1 strains, and modification levels were measured by HPLC. m5C levels for tRNAHis (black) and tRNAVal(AAC) (gray) during temperature-sensitive strain growth at 37°C were plotted. m5C values during growth at 24°C were plotted as the 37°C 0-h time point.

Melanie A. Preston, et al. RNA. 2013 February;19(2):243-256.
3.
FIGURE 4.

FIGURE 4. From: tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae.

Trm4-cMORF levels do not increase during starvation. BY4741 and Trm4-cMORF strains were grown in SD complete, SD – His, or SD – Leu media and grown for 24 h at 30°C. Crude extracts were prepared at 0, 7, and 24 h, and 30 μg of protein were loaded in each lane and resolved by SDS-PAGE, followed by Western blotting. Anti-HA antibody was used to detect the cMORF tag on Trm4. BY4741 crude extract was used as a negative control. The same blot was incubated with anti-enolase antibody as a loading control.

Melanie A. Preston, et al. RNA. 2013 February;19(2):243-256.
4.
FIGURE 5.

FIGURE 5. From: tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae.

Analysis of tRNAHis aminoacylation under different starvation conditions. RNA from the wild-type (BY4741) strain grown in the indicated media was isolated in acidic conditions to preserve aminoacylation, and 2 μg of RNA were resolved on an acidic gel, analyzed by Northern blot for tRNAHis, tRNAArg(ICG), and 5S rRNA, and quantified using ImageQuant software. A control sample was treated with base (+) to detect migration of deacylated tRNA. (Solid arrows) Aminoacyl-tRNA, (dotted arrows) deacyl-tRNA. Each percent aminoacylation value is the mean ± standard deviation (SD) from two independent cultures. A representative Northern blot from one of the two independent cultures is shown.

Melanie A. Preston, et al. RNA. 2013 February;19(2):243-256.
5.
FIGURE 1.

FIGURE 1. From: tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae.

Post-transcriptional modifications of mature tRNAHis. (A) Secondary structure of mature S. cerevisiae tRNAHis. Post-transcriptional modifications are indicated. G−1 is added to the 5′ end of tRNAHis by tRNAHis guanylyltransferase (Thg1). Under normal growth conditions, Trm4 methylates position 49 to form 5-methylcytidine (m5C, circle). When Thg1 is depleted, Trm4 also methylates adjacent cytidines at positions 48 and 50 (squares). (B) Representative HPLC traces of tRNAHis nucleosides. Wild-type (BY4741) and trm4-Δ cells were grown to log phase in SD complete media, and tRNAHis was purified, digested with P1 nuclease, phosphatase-treated, and separated by reverse-phase HPLC. Insets compare the wild type vs. trm4-Δ m5C peak (left), with the Am peak (right) as a control.

Melanie A. Preston, et al. RNA. 2013 February;19(2):243-256.
6.
FIGURE 3.

FIGURE 3. From: tRNAHis 5-methylcytidine levels increase in response to several growth arrest conditions in Saccharomyces cerevisiae.

tRNAHis m5C levels increase when BY4741 is grown to stationary phase. tRNAHis was purified from BY4741 grown in SD complete media in mid-log phase and in stationary phase for the indicated number of days. Nucleoside modifications were analyzed by HPLC, and m5C (black), Am (dark gray), and m1G + Gm (light gray) values are plotted over time. Other control modifications (dihydrouridine and pseudouridine) were unchanged during the time course (data not shown). The increase in m5C levels between log phase growth and Day 1 of stationary phase is highly reproducible (Day 0 log phase, m5C = 0.86 ± 0.03, n = 6; Day 1 stationary phase, m5C = 1.53 ± 0.11, n = 7, mean ± standard deviation) (see Table 3).

Melanie A. Preston, et al. RNA. 2013 February;19(2):243-256.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk