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Telomerase reverse transcriptase regulates microRNAs.

Lassmann T, Maida Y, Tomaru Y, Yasukawa M, Ando Y, Kojima M, Kasim V, Simon C, Daub CO, Carninci P, Hayashizaki Y, Masutomi K - Int J Mol Sci (2015)

Bottom Line: In humans, most microRNAs are transcribed by RNA polymerase II as long primary transcripts and processed by sequential cleavage of the two RNase III enzymes, DROSHA and DICER, into precursor and mature microRNAs, respectively.Although the fundamental functions of microRNAs in RNA silencing have been gradually uncovered, less is known about the regulatory mechanisms of microRNA expression.These results suggest that TERT regulates microRNAs at the very early phases in their biogenesis, presumably through non-telomerase mechanism(s).

View Article: PubMed Central - PubMed

Affiliation: RIKEN Omics Science Center, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan. lassmann@gsc.riken.jp.

ABSTRACT
MicroRNAs are small non-coding RNAs that inhibit the translation of target mRNAs. In humans, most microRNAs are transcribed by RNA polymerase II as long primary transcripts and processed by sequential cleavage of the two RNase III enzymes, DROSHA and DICER, into precursor and mature microRNAs, respectively. Although the fundamental functions of microRNAs in RNA silencing have been gradually uncovered, less is known about the regulatory mechanisms of microRNA expression. Here, we report that telomerase reverse transcriptase (TERT) extensively affects the expression levels of mature microRNAs. Deep sequencing-based screens of short RNA populations revealed that the suppression of TERT resulted in the downregulation of microRNAs expressed in THP-1 cells and HeLa cells. Primary and precursor microRNA levels were also reduced under the suppression of TERT. Similar results were obtained with the suppression of either BRG1 (also called SMARCA4) or nucleostemin, which are proteins interacting with TERT and functioning beyond telomeres. These results suggest that TERT regulates microRNAs at the very early phases in their biogenesis, presumably through non-telomerase mechanism(s).

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Related in: MedlinePlus

Mature miRNAs and their precursors are downregulated upon TERT suppression. The expression levels of primary, precursor and mature miRNAs in HeLa cells were analyzed with RT-qPCR under the suppression of TERT by sh-TERT#1 (gray) or sh-TERT#2 (black). The relative amount of the individual miRNAs was normalized to U6, and the mean expression levels of normalized sh-GFP (white) refer to one. (a,b) RT-qPCR of mature (a) and primary (b) miRNAs performed with total RNAs. The mRNA levels of ACTB and GAPDH were also examined with the same total RNAs; (c) RNAs smaller than 200 nt were used for RT-qPCR of mature (left) and precursor (right) miRNAs. Values represent the means ± SD for three independent experiments. Asterisks represent p < 0.05 compared with sh-GFP.
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ijms-16-01192-f002: Mature miRNAs and their precursors are downregulated upon TERT suppression. The expression levels of primary, precursor and mature miRNAs in HeLa cells were analyzed with RT-qPCR under the suppression of TERT by sh-TERT#1 (gray) or sh-TERT#2 (black). The relative amount of the individual miRNAs was normalized to U6, and the mean expression levels of normalized sh-GFP (white) refer to one. (a,b) RT-qPCR of mature (a) and primary (b) miRNAs performed with total RNAs. The mRNA levels of ACTB and GAPDH were also examined with the same total RNAs; (c) RNAs smaller than 200 nt were used for RT-qPCR of mature (left) and precursor (right) miRNAs. Values represent the means ± SD for three independent experiments. Asterisks represent p < 0.05 compared with sh-GFP.

Mentions: To further verify the deep sequencing findings, we quantified the expression levels of selected mature miRNAs under TERT suppression using quantitative RT-PCR (RT-qPCR). Corresponding with the deep sequencing findings (Figure 1), the RT-qPCR results indicated that mature miRNAs were apparently downregulated in both HeLa cells and THP-1 cells after reduction of TERT, irrespective of the suppression methods (Figure 2a and Figure S8). Taken together, these data confirm that TERT regulates mature miRNA levels in human cells.


Telomerase reverse transcriptase regulates microRNAs.

Lassmann T, Maida Y, Tomaru Y, Yasukawa M, Ando Y, Kojima M, Kasim V, Simon C, Daub CO, Carninci P, Hayashizaki Y, Masutomi K - Int J Mol Sci (2015)

Mature miRNAs and their precursors are downregulated upon TERT suppression. The expression levels of primary, precursor and mature miRNAs in HeLa cells were analyzed with RT-qPCR under the suppression of TERT by sh-TERT#1 (gray) or sh-TERT#2 (black). The relative amount of the individual miRNAs was normalized to U6, and the mean expression levels of normalized sh-GFP (white) refer to one. (a,b) RT-qPCR of mature (a) and primary (b) miRNAs performed with total RNAs. The mRNA levels of ACTB and GAPDH were also examined with the same total RNAs; (c) RNAs smaller than 200 nt were used for RT-qPCR of mature (left) and precursor (right) miRNAs. Values represent the means ± SD for three independent experiments. Asterisks represent p < 0.05 compared with sh-GFP.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307298&req=5

ijms-16-01192-f002: Mature miRNAs and their precursors are downregulated upon TERT suppression. The expression levels of primary, precursor and mature miRNAs in HeLa cells were analyzed with RT-qPCR under the suppression of TERT by sh-TERT#1 (gray) or sh-TERT#2 (black). The relative amount of the individual miRNAs was normalized to U6, and the mean expression levels of normalized sh-GFP (white) refer to one. (a,b) RT-qPCR of mature (a) and primary (b) miRNAs performed with total RNAs. The mRNA levels of ACTB and GAPDH were also examined with the same total RNAs; (c) RNAs smaller than 200 nt were used for RT-qPCR of mature (left) and precursor (right) miRNAs. Values represent the means ± SD for three independent experiments. Asterisks represent p < 0.05 compared with sh-GFP.
Mentions: To further verify the deep sequencing findings, we quantified the expression levels of selected mature miRNAs under TERT suppression using quantitative RT-PCR (RT-qPCR). Corresponding with the deep sequencing findings (Figure 1), the RT-qPCR results indicated that mature miRNAs were apparently downregulated in both HeLa cells and THP-1 cells after reduction of TERT, irrespective of the suppression methods (Figure 2a and Figure S8). Taken together, these data confirm that TERT regulates mature miRNA levels in human cells.

Bottom Line: In humans, most microRNAs are transcribed by RNA polymerase II as long primary transcripts and processed by sequential cleavage of the two RNase III enzymes, DROSHA and DICER, into precursor and mature microRNAs, respectively.Although the fundamental functions of microRNAs in RNA silencing have been gradually uncovered, less is known about the regulatory mechanisms of microRNA expression.These results suggest that TERT regulates microRNAs at the very early phases in their biogenesis, presumably through non-telomerase mechanism(s).

View Article: PubMed Central - PubMed

Affiliation: RIKEN Omics Science Center, RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan. lassmann@gsc.riken.jp.

ABSTRACT
MicroRNAs are small non-coding RNAs that inhibit the translation of target mRNAs. In humans, most microRNAs are transcribed by RNA polymerase II as long primary transcripts and processed by sequential cleavage of the two RNase III enzymes, DROSHA and DICER, into precursor and mature microRNAs, respectively. Although the fundamental functions of microRNAs in RNA silencing have been gradually uncovered, less is known about the regulatory mechanisms of microRNA expression. Here, we report that telomerase reverse transcriptase (TERT) extensively affects the expression levels of mature microRNAs. Deep sequencing-based screens of short RNA populations revealed that the suppression of TERT resulted in the downregulation of microRNAs expressed in THP-1 cells and HeLa cells. Primary and precursor microRNA levels were also reduced under the suppression of TERT. Similar results were obtained with the suppression of either BRG1 (also called SMARCA4) or nucleostemin, which are proteins interacting with TERT and functioning beyond telomeres. These results suggest that TERT regulates microRNAs at the very early phases in their biogenesis, presumably through non-telomerase mechanism(s).

Show MeSH
Related in: MedlinePlus