title: Quantifying mRNA targeting to P-bodies in living human cells reveals their dual role in mRNA decay and storage.
abstract: The 5'-to-3' mRNA degradation machinery localizes to cytoplasmic processing bodies (P-bodies), which are non-membranous structures found in all eukaryotes. Although P-body function has been intensively studied in yeast, less is known about their role in mammalian cells, such as whether P-body enzymes are actively engaged in mRNA degradation or whether P-bodies serve as mRNA storage depots, particularly during cellular stress. We examined the fate of mammalian mRNAs in P-bodies during translational stress, and show that mRNAs accumulate within P-bodies during amino acid starvation. The 5' and 3' ends of the transcripts residing in P-bodies could be identified, but poly(A) tails were not detected. Using the MS2 mRNA-tagging system for mRNA visualization in living cells, we found that a stationary mRNA population formed in P-bodies during translational stress, which cleared gradually after the stress was relieved. Dcp2-knockdown experiments showed that there is constant degradation of part of the P-body-associated mRNA population. This analysis demonstrates the dual role of P-bodies as decay sites and storage areas under regular and stress conditions.
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Adva
authors.initials: A
authors.lastname: Aizer
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Alon
authors.initials: A
authors.lastname: Kalo
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Pinhas
authors.initials: P
authors.lastname: Kafri
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Amit
authors.initials: A
authors.lastname: Shraga
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Rakefet
authors.initials: R
authors.lastname: Ben-Yishay
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Avi
authors.initials: A
authors.lastname: Jacob
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.
authors.forename: Noa
authors.initials: N
authors.lastname: Kinor
authors.affiliation: The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel Yaron.Shav-Tal@biu.ac.il.
authors.forename: Yaron
authors.initials: Y
authors.lastname: Shav-Tal
mesh_terms: D000596:Amino Acids / Q000172:deficiency; D002460:Cell Line; D022082:Cellular Structures / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D014176:Protein Biosynthesis; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics*; D013312:Stress, Physiological / Q000235:genetics; D059008:Time-Lapse Imaging
chemical_list: D000596:Amino Acids; D012333:RNA, Messenger; D004722:Endoribonucleases; C467723:DCP2 protein, human
keywords: P-body; RNA dynamics; RNA quantification