Abstract
Imaging of the microtubular network is an important strategy to define cell-cycle specific and pathological states of eukaryotic cells. Here, we describe TubStain, a novel recombinant polypeptide allowing direct, non-antibody dependent labeling of microtubules in fixed cells over a broad range of different species and tissues. TubStain has, due to its small size and susceptibility for biochemical and genetic manipulations, high potential as a microtubule marker in state-of-the-art microscopy.
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References
Bharti P, Schliebs W, Schievelbusch T, Neuhaus A, David C, Kock K, Herrmann C, Meyer HE, Wiese S, Warscheid B, Theiss C, Erdmann R (2011) PEX14 is required for microtubule-based peroxisome motility in human cells. J Cell Sci 124(Pt 10):1759–1768
Conde C, Caceres A (2009) Microtubule assembly, organization and dynamics in axons and dendrites. Nat Rev Neurosci 10(5):319–332
Farre JC, Krick R, Subramani S, Thumm M (2009) Turnover of organelles by autophagy in yeast. Curr Opin Cell Biol 21(4):522–530
Fernandez-Suarez M, Ting AY (2008) Fluorescent probes for super-resolution imaging in living cells. Nat Rev Mol Cell Biol 9(12):929–943
Giessmann D, Theiss C, Breipohl W, Meller K (2005) Decreased gap junctional communication in neurobiotin microinjected lens epithelial cells after taxol treatment. Anat Embryol (Berl) 209(5):391–400
Hara-Kuge S, Fujiki Y (2008) The peroxin Pex14p is involved in LC3-dependent degradation of mammalian peroxisomes. Exp Cell Res 314(19):3531–3541
Itoh R, Fujiki Y (2006) Functional domains and dynamic assembly of the peroxin Pex14p, the entry site of matrix proteins. J Biol Chem 281(15):10196–10205
Janke C, Bulinski JC (2011) Post-translational regulation of the microtubule cytoskeleton: mechanisms and functions. Nat Rev Mol Cell Biol 12(12):773–786
Kamath K, Oroudjev E, Jordan MA (2010) Determination of microtubule dynamic instability in living cells. Methods Cell Biol 97:1–14
Krah K, Meller K (1999) Axonal and dendritic transport in Purkinje cells of cerebellar slice cultures studied by microinjection of horseradish peroxidase. Cell Tissue Res 295(1):55–64
Meinecke M, Cizmowski C, Schliebs W, Kruger V, Beck S, Wagner R, Erdmann R (2010) The peroxisomal importomer constitutes a large and highly dynamic pore. Nat Cell Biol 12(3):273–277
Neufeld C, Filipp FV, Simon B, Neuhaus A, Schuller N, David C, Kooshapur H, Madl T, Erdmann R, Schliebs W, Wilmanns M, Sattler M (2009) Structural basis for competitive interactions of Pex14 with the import receptors Pex5 and Pex19. EMBO J 28(6):745–754
Parekh H, Simpkins H (1997) The transport and binding of taxol. Gen Pharmacol 29(2):167–172
Pfleger BF, Fawzi NJ, Keasling JD (2005) Optimization of DsRed production in Escherichia coli: effect of ribosome binding site sequestration on translation efficiency. Biotechnol Bioeng 92(5):553–558
Ravelli RB, Gigant B, Curmi PA, Jourdain I, Lachkar S, Sobel A, Knossow M (2004) Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain. Nature 428(6979):198–202
Risinger AL, Giles FJ, Mooberry SL (2009) Microtubule dynamics as a target in oncology. Cancer Treat Rev 35(3):255–261
Schiff PB, Horwitz SB (1980) Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci USA 77(3):1561–1565
Stanley WA, Filipp FV, Kursula P, Schuller N, Erdmann R, Schliebs W, Sattler M, Wilmanns M (2006) Recognition of a functional peroxisome type 1 target by the dynamic import receptor Pex5p. Mol Cell 24(5):653–663
Stanton RA, Gernert KM, Nettles JH, Aneja R (2011) Drugs that target dynamic microtubules: a new molecular perspective. Med Res Rev 31(3):443–481
Su JR, Takeda K, Tamura S, Fujiki Y, Miki K (2009) Crystal structure of the conserved N-terminal domain of the peroxisomal matrix protein import receptor, Pex14p. Proc Natl Acad Sci USA 106(2):417–421
Su JR, Takeda K, Tamura S, Fujiki Y, Miki K (2010) Monomer-dimer transition of the conserved N-terminal domain of the mammalian peroxisomal matrix protein import receptor, Pex14p. Biochem Biophys Res Commun 394(1):217–221
Theiss C, Meller K (2000) Taxol impairs anterograde axonal transport of microinjected horseradish peroxidase in dorsal root ganglia neurons in vitro. Cell Tissue Res 299(2):213–224
Theiss C, Meller K (2002) Microinjected anti-actin antibodies decrease gap junctional intercellular commmunication in cultured astrocytes. Exp Cell Res 281(2):197–204
Theiss C, Napirei M, Meller K (2005) Impairment of anterograde and retrograde neurofilament transport after anti-kinesin and anti-dynein antibody microinjection in chicken dorsal root ganglia. Eur J Cell Biol 84(1):29–43
Unger T, Jacobovitch Y, Dantes A, Bernheim R, Peleg Y (2010) Applications of the Restriction Free (RF) cloning procedure for molecular manipulations and protein expression. J Struct Biol 172(1):34–44
Zacharias DA, Violin JD, Newton AC, Tsien RY (2002) Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells. Science 296(5569):913–916
Zutphen T, Veenhuis M, van der Klei IJ (2008) Pex14 is the sole component of the peroxisomal translocon that is required for pexophagy. Autophagy 4(1):63–66
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C. Theiss and A. Neuhaus contributed equally to this work.
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Theiss, C., Neuhaus, A., Schliebs, W. et al. TubStain: a universal peptide-tool to label microtubules. Histochem Cell Biol 138, 531–540 (2012). https://doi.org/10.1007/s00418-012-0992-6
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DOI: https://doi.org/10.1007/s00418-012-0992-6