Molecular architecture of the rod domain of the Dictyostelium gelation factor (ABP120)

Fucini, Paola; Köppel, Bernd; Schleicher, Michael; Lustig, Ariel; Holak, Tad A.; Müller, Rolf; Stewart, Murray; Noegel, Angelika A.

doi:10.1006/jmbi.1999.3046

Cited by 28 publications

(19 citation statements)

References 27 publications

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“…The constructs were generated by PCR and verified by sequencing. In all full-length proteins a linker of four glycine residues separated the GFP moiety from the C-terminal rod repeat to ensure dimerization, which occurs via the last repeat, repeat 6 (25,29).…”

Section: Methodsmentioning

confidence: 99%

“…Filamins dimerize through the last repeats that interact with each other to form an extended ␤-sheet (25,29). The very last two C-terminal residues in those repeats are preceded by an 8-residue ␤-strand, both amino acid stretches are far away from the dimerization interface, and the placement of a tag at this end of the protein should not interfere with dimerization.…”

Section: Properties Of Full-length and Truncated Filamin Proteins-inmentioning

confidence: 99%

“…Structurally filamins are homodimers with large polypeptide chains that associate at their C termini (24,25). Their conserved N-terminal actin-binding domains (ABD) 2 belong to the ABDs common to the members of the ␣-actinin/spectrin superfamily of actin-binding proteins (26,27).…”

mentioning

confidence: 99%

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Filamin-regulated F-actin Assembly Is Essential for Morphogenesis and Controls Phototaxis in Dictyostelium

Khaire

Müller

Blau-Wasser

et al. 2007

Journal of Biological Chemistry

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Dictyostelium strains lacking the F-actin cross-linking protein filamin (ddFLN) have a severe phototaxis defect at the multicellular slug stage. Filamins are rod-shaped homodimers that cross-link the actin cytoskeleton into highly viscous, orthogonal networks. Each monomer chain of filamin is comprised of an F-actin-binding domain and a rod domain. In rescue experiments only intact filamin re-established correct phototaxis in filamin minus mutants, whereas C-terminally truncated filamin proteins that had lost the dimerization domain and molecules lacking internal repeats but retaining the dimerization domain did not rescue the phototaxis defect. Deletion of individual rod repeats also changed their subcellular localization, and mutant filamins in general were less enriched at the cell cortex as compared with the full-length protein and were increasingly present in the cytoplasm. For correct phototaxis ddFLN is only required at the tip of the slug because expression under control of the cell type-specific extracellular-matrix protein A (ecmA) promoter and mixing experiments with wild type cells supported phototactic orientation. Likewise, in chimeric slugs wild type cells were primarily found at the tip of the slug, which acts as an organizer in Dictyostelium morphogenesis.

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Section: Methodsmentioning

confidence: 99%

Section: Properties Of Full-length and Truncated Filamin Proteins-inmentioning

confidence: 99%

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Filamin-regulated F-actin Assembly Is Essential for Morphogenesis and Controls Phototaxis in Dictyostelium

Khaire

Müller

Blau-Wasser

et al. 2007

Journal of Biological Chemistry

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“…The contour lengths L, measured by means of the WLC fits to the data, had to be calibrated to obtain the number of amino acids involved in an unfolding event. For calibration, we used Ig domains of DdFLN, which contain exactly 100 amino acids per domain (20). The average contour length increase after the unfolding of a single DdFLN domain is ⌬L ϭ 32.5 Ϯ 0.3 nm.…”

Section: Elastically Coupled Two-state Modelmentioning

confidence: 99%

Exploring the energy landscape of GFP by single-molecule mechanical experiments

Dietz

Rief

2004

Proc. Natl. Acad. Sci. U.S.A.

332

296

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“…These distinct structures can be attributed to specific activities of individual actin-crosslinking proteins or protein families like the filamins (Ayscough, 1998;Stossel et al, 2001). Structurally, filamins are dimers with large polypeptide chains that associate at their C-termini (Gorlin et al, 1990;Fucini et al, 1999). Their conserved N-terminal actin-binding domains resemble the ones of the α-actinin/spectrin superfamily of actin-binding proteins (Hartwig, 1994).…”

Section: Introductionmentioning

confidence: 99%

A novel partner forDictyosteliumfilamin is an α-helical developmentally regulated protein

Knuth

Khaire

Kuspa

et al. 2004

Journal of Cell Science

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The filamins are a family of highly homologous actincrosslinking proteins that stabilize three-dimensional actin networks, link them to membrane proteins and direct intracellular signaling reactions to the actin scaffold through interaction with various binding partners. Here, we describe the first Dictyostelium filamin-interacting protein to be isolated - FIP, a 229.8 kDa protein with two α-helical coiled coil domains. FIP was identified in a yeast two-hybrid screen using the rod domain of filamin as bait. FIP can also be coimmunoprecipitated with filamin from cellular extracts. Deletion analysis located the interaction domain of FIP to a C-terminal region; by contrast, in filamin rods, repeats 2-4 interacted with the recombinant FIP protein. The 7 kb transcript of FIP is upregulated during early development. Monoclonal antibodies raised against a bacterially expressed FIP polypeptide recognize a 230 kDa developmentally regulated protein in western blots. Immunofluorescence analysis shows a punctate staining pattern in the cytosol and, in cell fractionation experiments, FIP is mainly found in the cytosolic fraction. A fusion protein composed of GFP and the C-terminal part localizes to the plasma membrane and is associated with the cytoskeleton. Expression of the fusion protein affects development and influences the size of the multicellular aggregates and the phototactic behavior of slugs. Thus, FIP might provide a candidate link between the dynamic actin cytoskeleton and signal transduction events during the multicellular stages of Dictyostelium amoebae.

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Molecular architecture of the rod domain of the Dictyostelium gelation factor (ABP120)

Cited by 28 publications

References 27 publications

Filamin-regulated F-actin Assembly Is Essential for Morphogenesis and Controls Phototaxis in Dictyostelium

Filamin-regulated F-actin Assembly Is Essential for Morphogenesis and Controls Phototaxis in Dictyostelium

Exploring the energy landscape of GFP by single-molecule mechanical experiments

A novel partner forDictyosteliumfilamin is an α-helical developmentally regulated protein

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