Skip to main content
Log in

Changes in distribution of actin mRNA in different polysome fractions following stimulation of MPC-11 cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Individual mRNA species have been shown to differ both with respect to localization in the cell, and in their distribution upon stimulation of cells with different signals. In this study we have examined the distribution of actin mRNA in the free, cytoskeletal-bound, and membrane-bound RNA fractions, both in starved cells, and in response to stimulation by feeding. These results were then compared with mRNAs for glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and histone H4. The results we obtained showed that actin mRNA was located in the free RNA fraction in starved cells, while upon stimulation it was located both in the free, and in the cytoskeletal fraction; no redistribution of GAPDH mRNA occurred between the three RNA fractions, while H4 mRNA showed a different localization upon stimulation. Incubation with the drugs actinomycin-D and cycloheximide showed that an altered localization of actin mRNA from free in starved cells to free and cytoskeletal mRNA fractions following stimulation, was dependent on RNA synthesis, and not on protein synthesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Ainger K, Avossa D, Morgan F, Hill JS, Barry C, Barbarese E, Carson JH: Transport and localization of endogenous myelin basic protein mRNA microinjected into oligodendrocytes. J Cell Biol 123: 431–441, 1993

    PubMed  Google Scholar 

  2. Bassel GJ: High resolution distribution of mRNA within the cytoskeleton. J Cell Biochem 52: 127–133, 1993

    PubMed  Google Scholar 

  3. Berleth T, Burri M, Thoma G, Bopp D, Richstein S, Frigerio G, Noll M, Nusslein-Volhard C: The role of localization ofbicoid RNA in organizing the anterior pattern of theDrosophila embryo. EMBO J 7: 1749–1756, 1988

    PubMed  Google Scholar 

  4. Bonneau AM, Darveau A, Sonenberg N: Effect of viral infection on host protein synthesis and mRNA association with the cytoplasmic cytoskeletal structure. J Cell Biol 100: 1209–1218, 1985

    PubMed  Google Scholar 

  5. Brodeur RD, Jeffery WR: Translational control and the cytoskeleton in Physarum polycephalum. Cell Motil Cytoskeleton 7: 129–137, 1987

    PubMed  Google Scholar 

  6. Bruckenstein DA, Lein PJ, Higgins D, Fremeau RTJ: Distinct spatial localization of specific mRNAs in cultured sympathetic neurons. Neuron 5: 809–819, 1990

    PubMed  Google Scholar 

  7. Cheng H, Bjerknes M: Asymmetric distribution of actin mRNA and cytoskeletal pattern generation in polarized epithelial cells. J Mol Biol 210: 541–549, 1989

    PubMed  Google Scholar 

  8. Davis L, Banker GA, Steward O: Selective dendritic transport of RNA in hippocampal neurons in culture. Nature 330: 477–479, 1987

    PubMed  Google Scholar 

  9. Erickson PA, Feinstein SC, Lewis GP, Fisher SK: Glial fibrillary acidic protein and its mRNA: ultrastructural detection and determination of changes after CNS injury. J Struct Biol 108: 148–161, 1992

    PubMed  Google Scholar 

  10. Fontaine B, Sassoon D, Buckingham M, Changeux JP: Detection of the nicotinic acetylcholine receptor alpha-subunit mRNA byin situ hybridization at neuromuscular junctions of 15-day-old chick striated muscles. EMBO J 7: 603–609, 1988

    PubMed  Google Scholar 

  11. Garner CC, Tucker RP, Matus A: Selective localization of messenger RNA for cytoskeletal protein MAP2 in dendrites. Nature 336: 674–677, 1988

    PubMed  Google Scholar 

  12. Gottlieb E: Messenger RNA transport and localization. Curr Opin Cell Biol 2: 1080–1086, 1990

    PubMed  Google Scholar 

  13. Gottlieb E: The 3′ untraslated region of localized maternal messages contains a conserved motif in mRNA localization. Proc Natl Acad Sci USA 89: 7164–7168, 1992

    PubMed  Google Scholar 

  14. Gunning P, Ponte P, Okayama H, Engel J, Blau H, Kedes L: Isolation and characterization of full-length cDNA clones for human α-, β, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biochem 3: 787–795, 1983

    Google Scholar 

  15. Hesketh JE, Campbell GP, Piechaczyk M, Blanchard JM: Targeting of c-myc and β-globin coding sequences to cytoskeletal-bound polysomes by c-myc 3′ untranslated region. Biochem J 298: 143–148, 1994

    PubMed  Google Scholar 

  16. Hesketh JE, Campbell GP, Whitelaw PF:c-myc Messenger RNA in cytoskeletal-bound polysomes in fibroblasts. Biochem J 274: 607–609, 1991

    PubMed  Google Scholar 

  17. Hesketh JE, Pryme IF: Interaction between mRNA, ribosomes and the cytoskeleton. Biochem J 277: 1–10, 1991

    PubMed  Google Scholar 

  18. Hill MA, Gunning P: Beta and gamma actin mRNAs are differently located within myoblasts. J Cell Biol 122: 825–832, 1993

    PubMed  Google Scholar 

  19. Hoock TC, Newcomb PM, Herman IM: Beta actin and its mRNA are localized at the plasma membrane and the regions of moving cytoplasm during the cellular response to injury. J Cell Biol 112: 653–664, 1991

    PubMed  Google Scholar 

  20. Howell AM, Cool D, Hewitt J, Ydenberg B, Smith MJ, Honda BM: Organization and unusual expression of histone genes in the sea starPisaster ochraceus. J Mol Evol 25: 29–36, 1987

    Google Scholar 

  21. Jackson R: Cytoplasmic regulation of mRNA function: the importance of the 3′-untranslated region. Cell 74: 9–14, 1993

    PubMed  Google Scholar 

  22. Kislauskis EH, Li Z, Singer RH, Taneja KL: Isoform-specific 3′-untranslated sequences sort α-cardiac and β-cytoplasmic actin messenger RNAs to different cytoplasmic compartments. J Cell Biol 123: 165–172, 1993

    PubMed  Google Scholar 

  23. Kleiman R, Banker G, Steward O: Differential subcellular localization of particular mRNAs in hippocampal neurons in culture. Neuron 5: 821–830, 1990

    PubMed  Google Scholar 

  24. Lawrence JB, Singer RH: Intracellular localization of messenger RNAs for cytoskeletal proteins. Cell 45: 407–415, 1986

    PubMed  Google Scholar 

  25. Lawrence JB, Singer RH: Spatial organization of nucleic acid sequences within cells. Semin Cell Biol 2: 83–101, 1991

    PubMed  Google Scholar 

  26. Macdonald PM, Struhl G: cis-acting sequences responsible for anterior localization ofbicoid mRNA inDrosophila embryos. Nature 336: 595–598, 1988

    PubMed  Google Scholar 

  27. Masibay AS, Qasba PK, Sengupta DN, Damewood GP, Sreevalsan T: Cell-cycle-specific and serum-dependent expression of gamma-actin mRNA in Swiss mouse 3T3 cells. Mol Cell Biol 8: 2288–2294, 1988

    PubMed  Google Scholar 

  28. Messina JL, Weinstock RS: Regulation of β-actin gene transcription by insulin and phorbol esters. Exp Cell Res 200: 532–535, 1992

    PubMed  Google Scholar 

  29. Moss R, Pryme IF, Vedeler A: The effect of insulin on the proteins associated with free, cytoskeletal-bound and membrane-bound polysome populations. Cell Biol Int 17: 1065–1073, 1993

    PubMed  Google Scholar 

  30. Moss R, Pryme IF, Vedeler A: Free, cytoskeletal-bound and membrane-bound polysomes isolated from MPC-11 and Krebs II ascites cells differ in their complement of poly(A) binding proteins. Mol Cell Biochem 131: 131–139, 1994

    PubMed  Google Scholar 

  31. Perry BA, Capco DG: Spatial reorganization of actin, tubulin and histone mRNAs during meiotic maturation and fertilization inXenopus oocytes. Cell Differ Dev 25: 99–108, 1988

    PubMed  Google Scholar 

  32. Pokrywka NJ, Stephenson EC: Microtubules mediate the localization of bicoidRNA duringDrosophila oogenesis. Development 113: 55–66, 1991

    PubMed  Google Scholar 

  33. Pryme IF: On the attachment of ribosomes to heavy and light rough endoplasmic reticulum in eucaryotic cells in Abraham AK, Eikhom TS, Pryme IF (eds): Protein synthesis. Clifton, New Jersey. The Humana press, 1983, pp 117–129

    Google Scholar 

  34. Pryme IF, Johannessen AJ, Vedeler A: The sequenstration of mRNA in the cytoskeleton and other subcellular structures. Adv Mol Cell Biol 1994 (in press)

  35. Rabinovich YM, Kreinin MO: Evidence for non-uniform translation of individual polypeptides in rat liver mitochondria. Biochim Biophys Acta 1089: 193–196, 1991

    PubMed  Google Scholar 

  36. Sharpless K, Biegel D, Yang T, Pachter JS: Beta-actin mRNA-binding proteins associated with the cytoskeletal framework. Eur J Biochem 212: 217–225, 1993

    PubMed  Google Scholar 

  37. Sundell CL, Singer RH: Actin mRNA localizes in the absence of protein synthesis. J Cell Biol 111: 2397–2403, 1990

    PubMed  Google Scholar 

  38. Sundell CL, Singer RH: Requirement of microfilaments in sorting of actin messenger RNA. Science 253: 1275–1277, 1991

    PubMed  Google Scholar 

  39. Taneja KL, Lifshitz LM, Fay FS, Singer RH: Poly(A) RNA codistribution with microfilaments: evaluation byin situ hybridization and quantitative digital imaging microscopy. J Cell Biol 119: 1245–1260, 1992

    PubMed  Google Scholar 

  40. Tso JY, Sun X, Kao T, Reece S, Wu R: Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucl Acids Res 13: 2485–2502, 1985

    PubMed  Google Scholar 

  41. Vedeler A, Pryme IF, Hesketh JE: Insulin and step-up conditions cause redistribution of polysomes among free, cytoskeletal-bound and membrane-bound fractions in Krebs II ascites cells. Cell Biol Int Rep 14: 211–218, 1990

    PubMed  Google Scholar 

  42. Vedeler A, Pryme IF, Hesketh JE: The characterization of free, cytoskeletal and membrane-bound polysomes in Krebs II ascites and 3T3 cells. Mol Cell Biochem 100: 183–193, 1991

    PubMed  Google Scholar 

  43. Vedeler A, Pryme IF, Hesketh JE: The compartmentalization of polysomes into free, cytoskeletal-bound and membrane-bound populations. Biochem Soc Trans 19: 1108–1111, 1991

    PubMed  Google Scholar 

  44. Weeks DL, Melton DA: A maternal mRNA localized to the vegetal hemisphere inXenopus eggs codes for a growth factor related to TGF-beta. Cell 51: 861–867, 1987

    PubMed  Google Scholar 

  45. Willhelm JE, Vale RD: RNA on the move: The mRNA localization pathway. J Cell Biol 123: 269–274, 1993

    PubMed  Google Scholar 

  46. Yisraeli JK, Sokol S, Melton DA: A two-step model for the localization of maternal mRNA inXenopus oocytes: involvement of microtubules and microfilaments in the translocation and anchoring of Vgl mRNA. Development 108: 289–298, 1990

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Johannessen, A.J., Pryme, I.F. & Vedeler, A. Changes in distribution of actin mRNA in different polysome fractions following stimulation of MPC-11 cells. Mol Cell Biochem 142, 107–115 (1995). https://doi.org/10.1007/BF00928931

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00928931

Key Words

Navigation