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Drosophila Ana2 is a conserved centriole duplication factor.

Stevens NR, Dobbelaere J, Brunk K, Franz A, Raff JW - J. Cell Biol. (2010)

Bottom Line: Functional orthologues of all but SAS-5 have been found in other species.Asl is now known to be essential for centriole duplication in flies, but no equivalent protein has been found in worms.We propose that members of the SAS-5/Ana2/STIL family of proteins are key conserved components of the centriole duplication machinery.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Gurdon Institute, Cambridge, England CB2 1QN, UK.

ABSTRACT
In Caenorhabditis elegans, five proteins are required for centriole duplication: SPD-2, ZYG-1, SAS-5, SAS-6, and SAS-4. Functional orthologues of all but SAS-5 have been found in other species. In Drosophila melanogaster and humans, Sak/Plk4, DSas-6/hSas-6, and DSas-4/CPAP-orthologues of ZYG-1, SAS-6, and SAS-4, respectively-are required for centriole duplication. Strikingly, all three fly proteins can induce the de novo formation of centriole-like structures when overexpressed in unfertilized eggs. Here, we find that of eight candidate duplication factors identified in cultured fly cells, only two, Ana2 and Asterless (Asl), share this ability. Asl is now known to be essential for centriole duplication in flies, but no equivalent protein has been found in worms. We show that Ana2 is the likely functional orthologue of SAS-5 and that it is also related to the vertebrate STIL/SIL protein family that has been linked to microcephaly in humans. We propose that members of the SAS-5/Ana2/STIL family of proteins are key conserved components of the centriole duplication machinery.

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Ana2 is a centriole component with a unique asymmetric localization. (A) Centriole pair from a G2 primary spermatocyte expressing Ana2-GFP (green) stained for the centriole marker GTU88* (red). Ana2-GFP localizes to the proximal and distal centriole ends and also exhibits a unique asymmetric distribution, localizing preferentially along one centriole barrel, which can be identified as the daughter from the GTU88* staining (see main text). (B) Centriole pair from a G2 primary spermatocyte expressing GFP-Ana2 (green) and RFP-PACT (red). GFP-Ana2 localization is indistinguishable from Ana2-GFP. (C) Centriole pair from a primary spermatocyte at anaphase of meiosis I: the centrioles are beginning to separate. The cell is expressing RFP-PACT (red) and GFP-Ana2 (green), which is no longer obviously asymmetric. (D) Two basal bodies from spermatids expressing RFP-PACT (blue) and GFP–DSas-6 (green), and stained for Ana2 (red). GFP–DSas-6 and Ana2 colocalize at the proximal centriole-like structure, a nodule adjacent to the basal body marked by RFP-PACT. Bars, 2 µm.
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fig3: Ana2 is a centriole component with a unique asymmetric localization. (A) Centriole pair from a G2 primary spermatocyte expressing Ana2-GFP (green) stained for the centriole marker GTU88* (red). Ana2-GFP localizes to the proximal and distal centriole ends and also exhibits a unique asymmetric distribution, localizing preferentially along one centriole barrel, which can be identified as the daughter from the GTU88* staining (see main text). (B) Centriole pair from a G2 primary spermatocyte expressing GFP-Ana2 (green) and RFP-PACT (red). GFP-Ana2 localization is indistinguishable from Ana2-GFP. (C) Centriole pair from a primary spermatocyte at anaphase of meiosis I: the centrioles are beginning to separate. The cell is expressing RFP-PACT (red) and GFP-Ana2 (green), which is no longer obviously asymmetric. (D) Two basal bodies from spermatids expressing RFP-PACT (blue) and GFP–DSas-6 (green), and stained for Ana2 (red). GFP–DSas-6 and Ana2 colocalize at the proximal centriole-like structure, a nodule adjacent to the basal body marked by RFP-PACT. Bars, 2 µm.

Mentions: We next wanted to compare the localization of Ana2 with that of the other Drosophila centriole duplication factors. DSas-4–GFP, GFP–DSas-6, and GFP-Sak are all enriched at the proximal and distal ends of the large spermatocyte centrioles (Peel et al., 2007). We found that, likewise, Ana2-GFP localized preferentially to the proximal and distal centriole tips. Strikingly, however, Ana2-GFP (and GFP-Ana2) also exhibited a unique asymmetric distribution, consistently localizing preferentially along one centriole barrel (Fig. 3, A and B).


Drosophila Ana2 is a conserved centriole duplication factor.

Stevens NR, Dobbelaere J, Brunk K, Franz A, Raff JW - J. Cell Biol. (2010)

Ana2 is a centriole component with a unique asymmetric localization. (A) Centriole pair from a G2 primary spermatocyte expressing Ana2-GFP (green) stained for the centriole marker GTU88* (red). Ana2-GFP localizes to the proximal and distal centriole ends and also exhibits a unique asymmetric distribution, localizing preferentially along one centriole barrel, which can be identified as the daughter from the GTU88* staining (see main text). (B) Centriole pair from a G2 primary spermatocyte expressing GFP-Ana2 (green) and RFP-PACT (red). GFP-Ana2 localization is indistinguishable from Ana2-GFP. (C) Centriole pair from a primary spermatocyte at anaphase of meiosis I: the centrioles are beginning to separate. The cell is expressing RFP-PACT (red) and GFP-Ana2 (green), which is no longer obviously asymmetric. (D) Two basal bodies from spermatids expressing RFP-PACT (blue) and GFP–DSas-6 (green), and stained for Ana2 (red). GFP–DSas-6 and Ana2 colocalize at the proximal centriole-like structure, a nodule adjacent to the basal body marked by RFP-PACT. Bars, 2 µm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2819680&req=5

fig3: Ana2 is a centriole component with a unique asymmetric localization. (A) Centriole pair from a G2 primary spermatocyte expressing Ana2-GFP (green) stained for the centriole marker GTU88* (red). Ana2-GFP localizes to the proximal and distal centriole ends and also exhibits a unique asymmetric distribution, localizing preferentially along one centriole barrel, which can be identified as the daughter from the GTU88* staining (see main text). (B) Centriole pair from a G2 primary spermatocyte expressing GFP-Ana2 (green) and RFP-PACT (red). GFP-Ana2 localization is indistinguishable from Ana2-GFP. (C) Centriole pair from a primary spermatocyte at anaphase of meiosis I: the centrioles are beginning to separate. The cell is expressing RFP-PACT (red) and GFP-Ana2 (green), which is no longer obviously asymmetric. (D) Two basal bodies from spermatids expressing RFP-PACT (blue) and GFP–DSas-6 (green), and stained for Ana2 (red). GFP–DSas-6 and Ana2 colocalize at the proximal centriole-like structure, a nodule adjacent to the basal body marked by RFP-PACT. Bars, 2 µm.
Mentions: We next wanted to compare the localization of Ana2 with that of the other Drosophila centriole duplication factors. DSas-4–GFP, GFP–DSas-6, and GFP-Sak are all enriched at the proximal and distal ends of the large spermatocyte centrioles (Peel et al., 2007). We found that, likewise, Ana2-GFP localized preferentially to the proximal and distal centriole tips. Strikingly, however, Ana2-GFP (and GFP-Ana2) also exhibited a unique asymmetric distribution, consistently localizing preferentially along one centriole barrel (Fig. 3, A and B).

Bottom Line: Functional orthologues of all but SAS-5 have been found in other species.Asl is now known to be essential for centriole duplication in flies, but no equivalent protein has been found in worms.We propose that members of the SAS-5/Ana2/STIL family of proteins are key conserved components of the centriole duplication machinery.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Gurdon Institute, Cambridge, England CB2 1QN, UK.

ABSTRACT
In Caenorhabditis elegans, five proteins are required for centriole duplication: SPD-2, ZYG-1, SAS-5, SAS-6, and SAS-4. Functional orthologues of all but SAS-5 have been found in other species. In Drosophila melanogaster and humans, Sak/Plk4, DSas-6/hSas-6, and DSas-4/CPAP-orthologues of ZYG-1, SAS-6, and SAS-4, respectively-are required for centriole duplication. Strikingly, all three fly proteins can induce the de novo formation of centriole-like structures when overexpressed in unfertilized eggs. Here, we find that of eight candidate duplication factors identified in cultured fly cells, only two, Ana2 and Asterless (Asl), share this ability. Asl is now known to be essential for centriole duplication in flies, but no equivalent protein has been found in worms. We show that Ana2 is the likely functional orthologue of SAS-5 and that it is also related to the vertebrate STIL/SIL protein family that has been linked to microcephaly in humans. We propose that members of the SAS-5/Ana2/STIL family of proteins are key conserved components of the centriole duplication machinery.

Show MeSH
Related in: MedlinePlus