title: A brilliant monomeric red fluorescent protein to visualize cytoskeleton dynamics in Dictyostelium.
abstract: Red fluorescent proteins (RFPs) combined with GFP are attractive probes for double-fluorescence labeling of proteins in live cells. However, the application of these proteins is restrained by stable oligomer formation and by their weak fluorescence in vivo. Previous attempts to eliminate these problems by mutagenesis of RFP from Discosoma (DsRed) resulted in the monomeric mRFP1 and in the tetrameric RedStar RFP, which is distinguished by its enhanced fluorescence in vivo. Based on these mutations, we have generated an enhanced monomeric RFP, mRFPmars, and report its spectral properties. Together with green fluorescent labels, we used mRFPmars to visualize filamentous actin structures and microtubules in Dictyostelium cells. This enhanced RFP proved to be suitable to monitor the dynamics of cytoskeletal proteins in cell motility, mitosis, and endocytosis using dual-wavelength fluorescence microscopy.
authors.affiliation: Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Fischer
authors.forename: Ilka
authors.initials: I
authors.lastname: Haase
authors.forename: Evelyn
authors.initials: E
authors.lastname: Simmeth
authors.forename: Günther
authors.initials: G
authors.lastname: Gerisch
authors.forename: Annette
authors.initials: A
authors.lastname: Müller-Taubenberger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003599:Cytoskeleton / Q000648:ultrastructure*; D004023:Dictyostelium / Q000648:ultrastructure*; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D017404:In Situ Hybridization, Fluorescence; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein