A conserved sequence in calmodulin regulated spectrin‐associated protein 1 links its interaction with spectrin and calmodulin to neurite outgrowth

King, Mikayala D.A.; Phillips, Gareth W.; Bignone, Paola A.; Hayes, Nandini V. L.; Pinder, Jennifer C.; Baines, Anthony J.

doi:10.1111/jnc.12462

Cited by 26 publications

(30 citation statements)

References 43 publications

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“…The highest scoring hit in this screen was the spectraplakin ACF7 ( Fig. 2A), whereas no peptides that were derived from spectrin, which has been previously reported to bind to CAMSAP1 (King et al, 2014), were found. Whether this was owing to the absence of appropriate spectrin isoforms in HEK293T cells, the biochemical procedure used or the lack of interaction between CAMSAP3 and spectrin is unclear.…”

Section: Acf7 Binds To Camsap3 and Participates In Its Apical Recruitmentioning

confidence: 88%

“…Experiments in Caco-2 cells, a human intestinal cancer cell line, have demonstrated that the first coiled-coil region of CAMSAP3 is involved in recruitment of CAMSAP3-bound microtubule minus ends to the apical side (Toya et al, 2016). In CAMSAP1, this coiled-coil region was shown to bind to spectrin (King et al, 2014). Whether the apical localization of CAMSAP3, indeed, depends on spectrin or some other proteins has not been determined.…”

Section: Introductionmentioning

confidence: 99%

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Control of apico–basal epithelial polarity by the microtubule minus-end-binding protein CAMSAP3 and spectraplakin ACF7

Noordstra

Liu

Nijenhuis

et al. 2016

Journal of Cell Science

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The microtubule cytoskeleton regulates cell polarity by spatially organizing membrane trafficking and signaling processes. In epithelial cells, microtubules form parallel arrays aligned along the apico-basal axis, and recent work has demonstrated that the members of CAMSAP/Patronin family control apical tethering of microtubule minus ends. Here, we show that in mammalian intestinal epithelial cells, the spectraplakin ACF7 (also known as MACF1) specifically binds to CAMSAP3 and is required for the apical localization of CAMSAP3-decorated microtubule minus ends. Loss of ACF7 but not of CAMSAP3 or its homolog CAMSAP2 affected the formation of polarized epithelial cysts in three-dimensional cultures. In short-term epithelial polarization assays, knockout of CAMSAP3, but not of CAMSAP2, caused microtubule re-organization into a more radial centrosomal array, redistribution of Rab11-positive (also known as Rab11A) endosomes from the apical cell surface to the pericentrosomal region and inhibition of actin brush border formation at the apical side of the cell. We conclude that ACF7 is an important regulator of apico-basal polarity in mammalian intestinal cells and that a radial centrosome-centered microtubule organization can act as an inhibitor of epithelial polarity.

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Section: Acf7 Binds To Camsap3 and Participates In Its Apical Recruitmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Control of apico–basal epithelial polarity by the microtubule minus-end-binding protein CAMSAP3 and spectraplakin ACF7

Noordstra

Liu

Nijenhuis

et al. 2016

Journal of Cell Science

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“…Although this result may reveal a difference in the biological function of Drosophila vs. mammalian proteins, it is possible that the knockdown of all three CAMSAPs may be needed to uncover a mitotic phenotype. CAMSAPs also have been reported to interact biochemically with spectrins (an activity from which the CAMSAPs derive their name), and overexpression studies have suggested that this interaction may be important in vivo (27). However, evidence supporting a physiological connection of this protein family with spectrin is less clear than their role in regulating MT minus-ends.…”

Section: Discussionmentioning

confidence: 99%

“…In mammals, Takeichi and coworkers (24,25) have shown that a homologous protein, which they termed Nezha, anchors minus-ends of MTs in cell-cell adherens junctions. The protein family was also identified through a spectrin binding activity and named calmodulin-regulated spectrin-associated protein (CAMSAP) (26,27). Bioinformatic analyses suggest that this protein family first evolved in metazoans; invertebrates possess a single gene whereas vertebrates possess three CAMSAP genes (CAMSAP1, CAMSAP2, and CAMSAP3/Nezha; Fig.…”

mentioning

confidence: 99%

Regulation of microtubule minus-end dynamics by CAMSAPs and Patronin

Hendershott

Vale

2014

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

159

184

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The microtubule (MT) cytoskeleton plays an essential role in mitosis, intracellular transport, cell shape, and cell migration. The assembly and disassembly of MTs, which can occur through the addition or loss of subunits at the plus-or minus-ends of the polymer, is essential for MTs to carry out their biological functions. A variety of proteins act on MT ends to regulate their dynamics, including a recently described family of MT minus-end binding proteins called calmodulin-regulated spectrin-associated protein (CAMSAP)/Patronin/Nezha. Patronin, the single member of this family in Drosophila, was previously shown to stabilize MT minusends against depolymerization in vitro and in vivo. Here, we show that all three mammalian CAMSAP family members also bind specifically to MT minus-ends and protect them against kinesin-13-induced depolymerization. However, these proteins differ in their abilities to suppress tubulin addition at minus-ends and to dissociate from MTs. CAMSAP1 does not interfere with polymerization and tracks along growing minus-ends. CAMSAP2 and CAMSAP3 decrease the rate of tubulin incorporation and remain bound, thereby creating stretches of decorated MT minus-ends. By using truncation analysis, we find that somewhat different minimal domains of CAMSAP and Patronin are involved in minus-end localization. However, we find that, in both cases, a highly conserved C-terminal domain and a more variable central domain cooperate to suppress minus-end dynamics in vitro and that both regions are required to stabilize minus-ends in Drosophila S2 cells. These results show that members of the CAMSAP/Patronin family all localize to and protect minus-ends but have evolved distinct effects on MT dynamics.tubulin polymerization | cytoskeletal regulation | TIRF microscopy M icrotubules (MTs) are cellular polymers that are important for a variety of functions, including cargo transport and mitotic spindle formation. MTs are composed of dimers of α-and β-tubulin that assemble head-to-tail, creating a polar protofilament. Protofilaments then assemble laterally to form the canonical 13-protofilament MT structure, with β-tubulin exposed at fast-growing plus-ends and α-tubulin exposed at slow-growing minus-ends (1). MTs exhibit an intriguing property termed "dynamic instability," whereby the polymer can abruptly switch between episodes of net growth and shrinkage (2). The rates of growth and shrinkage as well as the frequency of transitions between these two states are regulated by numerous MT-associated proteins, many of which bind to the ends of the polymer (3, 4).The dynamics of MT plus-ends are regulated by a well-characterized network of plus-end tracking proteins (+TIPs) (5). End-binding proteins recognize a tubulin conformation unique to the growing ends of MTs and can affect the dynamics of plusends by intrinsically altering the structure of the MT end (6-8) as well as recruiting other interacting proteins (9). In contrast, TOG domain-containing proteins, such as XMAP215, promote MT growth and have been suggested ...

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“…The CC1 domain of CAMSAP1 has been reported to interact with βII-spectrin (25). The amino acid sequence crucial for this interaction within the CC1 domain was identified as LEEK (25), and this sequence with additional R (i.e., LEEKR) is conserved in CAMSAP3 (Fig.…”

Section: Significancementioning

confidence: 99%

CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells

Toya¹,

Kobayashi²,

Kawasaki³

et al. 2015

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

129

171

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Polarized epithelial cells exhibit a characteristic array of microtubules that are oriented along the apicobasal axis of the cells. The minus-ends of these microtubules face apically, and the plus-ends face toward the basal side. The mechanisms underlying this epithelial-specific microtubule assembly remain unresolved, however. Here, using mouse intestinal cells and human Caco-2 cells, we show that the microtubule minus-end binding protein CAMSAP3 (calmodulin-regulated–spectrin-associated protein 3) plays a pivotal role in orienting the apical-to-basal polarity of microtubules in epithelial cells. In these cells, CAMSAP3 accumulated at the apical cortices, and tethered the longitudinal microtubules to these sites. Camsap3 mutation or depletion resulted in a random orientation of these microtubules; concomitantly, the stereotypic positioning of the nucleus and Golgi apparatus was perturbed. In contrast, the integrity of the plasma membrane was hardly affected, although its structural stability was decreased. Further analysis revealed that the CC1 domain of CAMSAP3 is crucial for its apical localization, and that forced mislocalization of CAMSAP3 disturbs the epithelial architecture. These findings demonstrate that apically localized CAMSAP3 determines the proper orientation of microtubules, and in turn that of organelles, in mature mammalian epithelial cells.

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A conserved sequence in calmodulin regulated spectrin‐associated protein 1 links its interaction with spectrin and calmodulin to neurite outgrowth

Cited by 26 publications

References 43 publications

Control of apico–basal epithelial polarity by the microtubule minus-end-binding protein CAMSAP3 and spectraplakin ACF7

Control of apico–basal epithelial polarity by the microtubule minus-end-binding protein CAMSAP3 and spectraplakin ACF7

Regulation of microtubule minus-end dynamics by CAMSAPs and Patronin

CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells

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