TY - JOUR
T1 - Reassessment of the Role of TSC, mTORC1 and MicroRNAs in Amino Acids-Meditated Translational Control of TOP mRNAs
AU - Patursky-Polischuk, Ilona
AU - Kasir, Judith
AU - Miloslavski, Rachel
AU - Hayouka, Zvi
AU - Hausner-Hanochi, Mirit
AU - Stolovich-Rain, Miri
AU - Tsukerman, Pinchas
AU - Biton, Moshe
AU - Mudhasani, Rajini
AU - Jones, Stephen N.
AU - Meyuhas, Oded
N1 - We thank Robert Abraham for mTOR-rr and mTOR-rr-kd plasmids, Stuart Schreiber for Flag-mTOR plasmid, Tatsuya Maeda for plasmids encoding Flag-mTOR and hyperactive mTORSL1+IT (pYO293), Raymond Stallings for Psi/mir10 and Psi/mir10mut, Michal Gropp for the Sin18 plasmid, David Sabatini (HHMI/Whitehead Institute) and Nathanael Gray (Dana-Farber Cancer Institute) for Torin1. Author Contributions: Conceived and designed the experiments: IPP OM. Performed the experiments: IPP JK R. Miloslavski ZH MHH MSR PT MB. Analyzed the data: IPP JK R. Miloslavski ZH MHH MSR PT MB. Contributed reagents/materials/analysis tools: R. Mudhasani SJ. Wrote the paper: IPP OM
PY - 2014/10/22
Y1 - 2014/10/22
N2 - TOP mRNAs encode components of the translational apparatus, and repression of their translation comprises one mechanism, by which cells encountering amino acid deprivation downregulate the biosynthesis of the protein synthesis machinery. This mode of regulation involves TSC as knockout of TSC1 or TSC2 rescued TOP mRNAs translation in amino acid-starved cells. The involvement of mTOR in translational control of TOP mRNAs is demonstrated by the ability of constitutively active mTOR to relieve the translational repression of TOP mRNA upon amino acid deprivation. Consistently, knockdown of this kinase as well as its inhibition by pharmacological means blocked amino acid-induced translational activation of these mRNAs. The signaling of amino acids to TOP mRNAs involves RagB, as overexpression of active RagB derepressed the translation of these mRNAs in amino acid-starved cells. Nonetheless, knockdown of raptor or rictor failed to suppress translational activation of TOP mRNAs by amino acids, suggesting that mTORC1 or mTORC2 plays a minor, if any, role in this mode of regulation. Finally, miR10a has previously been suggested to positively regulate the translation of TOP mRNAs. However, we show here that titration of this microRNA failed to downregulate the basal translation efficiency of TOP mRNAs. Moreover, Drosha knockdown or Dicer knockout, which carries out the first and second processing steps in microRNAs biosynthesis, respectively, failed to block the translational activation of TOP mRNAs by amino acid or serum stimulation. Evidently, these results are questioning the positive role of microRNAs in this mode of regulation.
AB - TOP mRNAs encode components of the translational apparatus, and repression of their translation comprises one mechanism, by which cells encountering amino acid deprivation downregulate the biosynthesis of the protein synthesis machinery. This mode of regulation involves TSC as knockout of TSC1 or TSC2 rescued TOP mRNAs translation in amino acid-starved cells. The involvement of mTOR in translational control of TOP mRNAs is demonstrated by the ability of constitutively active mTOR to relieve the translational repression of TOP mRNA upon amino acid deprivation. Consistently, knockdown of this kinase as well as its inhibition by pharmacological means blocked amino acid-induced translational activation of these mRNAs. The signaling of amino acids to TOP mRNAs involves RagB, as overexpression of active RagB derepressed the translation of these mRNAs in amino acid-starved cells. Nonetheless, knockdown of raptor or rictor failed to suppress translational activation of TOP mRNAs by amino acids, suggesting that mTORC1 or mTORC2 plays a minor, if any, role in this mode of regulation. Finally, miR10a has previously been suggested to positively regulate the translation of TOP mRNAs. However, we show here that titration of this microRNA failed to downregulate the basal translation efficiency of TOP mRNAs. Moreover, Drosha knockdown or Dicer knockout, which carries out the first and second processing steps in microRNAs biosynthesis, respectively, failed to block the translational activation of TOP mRNAs by amino acid or serum stimulation. Evidently, these results are questioning the positive role of microRNAs in this mode of regulation.
UR - http://www.scopus.com/inward/record.url?scp=84908610762&partnerID=8YFLogxK
U2 - https://doi.org/10.1371/journal.pone.0109410
DO - https://doi.org/10.1371/journal.pone.0109410
M3 - مقالة
C2 - 25338081
SN - 1932-6203
VL - 9
JO - PLoS ONE
JF - PLoS ONE
IS - 10
M1 - e109410
ER -