DOI: 10.1091/mbc.e02-11-0755. Three-dimensional Organization of Basal Bodies from Wild-Type and δ-Tubulin Deletion Strains of<i>Chlamydomonas reinhardtii</i>

Three-dimensional Organization of Basal Bodies from Wild-Type and δ-Tubulin Deletion Strains ofChlamydomonas reinhardtii

10.1091/mbc.e02-11-0755

Crossref journal-article

American Society for Cell Biology (ASCB)

Molecular Biology of the Cell (1076)

Abstract

Improved methods of specimen preparation and dual-axis electron tomography have been used to study the structure and organization of basal bodies in the unicellular alga Chlamydomonas reinhardtii. Novel structures have been found in both wild type and strains with mutations that affect specific tubulin isoforms. Previous studies have shown that strains lacking δ-tubulin fail to assemble the C-tubule of the basal body. Tomographic reconstructions of basal bodies from the δ-tubulin deletion mutant uni3-1 have confirmed that basal bodies contain mostly doublet microtubules. Our methods now show that the stellate fibers, which are present only in the transition zone of wild-type cells, repeat within the core of uni3-1 basal bodies. The distal striated fiber is incomplete in this mutant, rootlet microtubules can be misplaced, and multiflagellate cells have been observed. A suppressor of uni3-1, designated tua2-6, contains a mutation in α-tubulin. tua2-6; uni3-1 cells build both flagella, yet they retain defects in basal body structure and in rootlet microtubule positioning. These data suggest that the presence of specific tubulin isoforms in Chlamydomonas directly affects the assembly and function of both basal bodies and basal body-associated structures.

Bibliography

OâToole, E. T., Giddings, T. H., McIntosh, J. R., & Dutcher, S. K. (2003). Three-dimensional Organization of Basal Bodies from Wild-Type and Î´-Tubulin Deletion Strains ofChlamydomonas reinhardtii. Molecular Biology of the Cell, 14(7), 2999â3012.

Authors 4

Eileen T. O'Toole (first)
Thomas H. Giddings (additional)
J. Richard McIntosh (additional)
Susan K. Dutcher (additional)

References 46 Referenced 135

Dates

Type	When
Created	22 years, 4 months ago (April 7, 2003, 10:17 p.m.)
Deposited	4 years, 2 months ago (June 6, 2021, 1:45 a.m.)
Indexed	1 month, 3 weeks ago (June 30, 2025, 9:25 a.m.)
Issued	22 years, 1 month ago (July 1, 2003)
Published	22 years, 1 month ago (July 1, 2003)
Published Print	22 years, 1 month ago (July 1, 2003)

Funders 0

None

BibTeX

@article{O_Toole_2003, title={Three-dimensional Organization of Basal Bodies from Wild-Type and δ-Tubulin Deletion Strains ofChlamydomonas reinhardtii}, volume={14}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e02-11-0755}, DOI={10.1091/mbc.e02-11-0755}, number={7}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={O’Toole, Eileen T. and Giddings, Thomas H. and McIntosh, J. Richard and Dutcher, Susan K.}, year={2003}, month=jul, pages={2999–3012} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        6,
        30
      ]
    ],
    "date-time": "2025-06-30T13:25:06Z",
    "timestamp": 1751289906504
  },
  "reference-count": 46,
  "publisher": "American Society for Cell Biology (ASCB)",
  "issue": "7",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2003,
        7
      ]
    ]
  },
  "abstract": "<jats:p>Improved methods of specimen preparation and dual-axis electron tomography have been used to study the structure and organization of basal bodies in the unicellular alga Chlamydomonas reinhardtii. Novel structures have been found in both wild type and strains with mutations that affect specific tubulin isoforms. Previous studies have shown that strains lacking \u03b4-tubulin fail to assemble the C-tubule of the basal body. Tomographic reconstructions of basal bodies from the \u03b4-tubulin deletion mutant uni3-1 have confirmed that basal bodies contain mostly doublet microtubules. Our methods now show that the stellate fibers, which are present only in the transition zone of wild-type cells, repeat within the core of uni3-1 basal bodies. The distal striated fiber is incomplete in this mutant, rootlet microtubules can be misplaced, and multiflagellate cells have been observed. A suppressor of uni3-1, designated tua2-6, contains a mutation in \u03b1-tubulin. tua2-6; uni3-1 cells build both flagella, yet they retain defects in basal body structure and in rootlet microtubule positioning. These data suggest that the presence of specific tubulin isoforms in Chlamydomonas directly affects the assembly and function of both basal bodies and basal body-associated structures.</jats:p>",
  "DOI": "10.1091/mbc.e02-11-0755",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2003,
        4,
        8
      ]
    ],
    "date-time": "2003-04-08T02:17:26Z",
    "timestamp": 1049768246000
  },
  "page": "2999-3012",
  "source": "Crossref",
  "is-referenced-by-count": 135,
  "title": "Three-dimensional Organization of Basal Bodies from Wild-Type and \u03b4-Tubulin Deletion Strains of<i>Chlamydomonas reinhardtii</i>",
  "prefix": "10.1091",
  "volume": "14",
  "author": [
    {
      "given": "Eileen T.",
      "family": "O'Toole",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Boulder Laboratory for 3-D Fine Structure, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309-0347"
        }
      ]
    },
    {
      "given": "Thomas H.",
      "family": "Giddings",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Boulder Laboratory for 3-D Fine Structure, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309-0347"
        }
      ]
    },
    {
      "given": "J. Richard",
      "family": "McIntosh",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Boulder Laboratory for 3-D Fine Structure, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309-0347"
        }
      ]
    },
    {
      "given": "Susan K.",
      "family": "Dutcher",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110"
        }
      ]
    }
  ],
  "member": "1076",
  "reference": [
    {
      "key": "REF1",
      "doi-asserted-by": "crossref",
      "unstructured": "Adams, G.M.W., Wright, R.L., and Jarvik, J.W. (1985). Defective temporal and spatial control of flagellar assembly in a mutant ofChlamydomonas reinhardtiiwith variable flagellar number.J. Cell Biol.100, 955-964.",
      "DOI": "10.1083/jcb.100.3.955"
    },
    {
      "key": "REF2",
      "doi-asserted-by": "crossref",
      "unstructured": "Bullitt, E., Rout, M.P., Kilmartin, J.V., and Akey, C.W. (1997). The yeast spindle pole body is assemble around a central crystal of Spc42p.Cell89, 1077-1086.",
      "DOI": "10.1016/S0092-8674(00)80295-0"
    },
    {
      "key": "REF3",
      "doi-asserted-by": "crossref",
      "unstructured": "Cavalier-Smith, T. (1974). Basal bodies and flagellar development during the vegetative cell cycle and the sexual cycle ofChlamydomonas reinhardtii.J. Cell Sci.16, 529-556.",
      "DOI": "10.1242/jcs.16.3.529"
    },
    {
      "key": "REF4",
      "doi-asserted-by": "crossref",
      "unstructured": "Chang, P., and Stearns, T. (2000). \u03b4-Tubulin and \u03f5-tubulin: two new human centrosomal tubulins reveal new aspects of centrosome structure and function.Nat. Cell Biol.2, 30-35.",
      "DOI": "10.1038/71350"
    },
    {
      "key": "REF5",
      "doi-asserted-by": "crossref",
      "unstructured": "Dippel, R.V. (1968). The development of basal bodies inParamecium.Proc. Natl. Acad. Sci. USA61, 461-468.",
      "DOI": "10.1073/pnas.61.2.461"
    },
    {
      "key": "REF6",
      "doi-asserted-by": "crossref",
      "unstructured": "Doxsey, S. (2001). Re-evaluating centrosome function.Nat. Rev. Mol. Cell. Biol.2, 688-698.",
      "DOI": "10.1038/35089575"
    },
    {
      "key": "REF7",
      "doi-asserted-by": "crossref",
      "unstructured": "Dutcher, S.K. (2001). The tubulin fraternity: alpha to eta.Curr. Opin. Cell Biol.13, 49-54.",
      "DOI": "10.1016/S0955-0674(00)00173-3"
    },
    {
      "key": "REF8",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.e02-04-0205"
    },
    {
      "key": "REF9",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.9.6.1293"
    },
    {
      "key": "REF10",
      "doi-asserted-by": "crossref",
      "unstructured": "Ehler, L.L., Holmes, J.A., and Dutcher, S.K. (1995). Loss of spatial control of the mitotic spindle apparatus in aChlamydomonas reinhardtiimutant strain lacking basal bodies.Genetics141, 945-960.",
      "DOI": "10.1093/genetics/141.3.945"
    },
    {
      "key": "REF11",
      "doi-asserted-by": "crossref",
      "unstructured": "Frank, J. (1992). Introduction: principles of electron tomography. In:Electron Tomography: Three Dimensional Imaging with the Transmission Electron Microscope, ed. J. Frank, New York: Plenum Press, 1-13.",
      "DOI": "10.1007/978-1-4757-2163-8_1"
    },
    {
      "key": "REF12",
      "doi-asserted-by": "crossref",
      "unstructured": "Garreau de Loubresse, N., Ruiz, F., Beisson, J., and Klotz, C. (2001). Role of delta-tubulin and the C-tubule in assembly ofParameciumbasal bodies.BMC Cell Biol.2, 4-10.",
      "DOI": "10.1186/1471-2121-2-4"
    },
    {
      "key": "REF13",
      "unstructured": "Goodenough, U. W., and St. Clair, H. S. (1975). BALD-2: a mutation affecting the formation of doublet and triplet sets of MTs inChlamydomonas reinhardtii.J. Cell Biol.66: 480-491."
    },
    {
      "key": "REF14",
      "doi-asserted-by": "crossref",
      "unstructured": "Holmes, J.A., and Dutcher, S.K. (1989). Cellular asymmetry inChlamydomonas reinhardtii.J. Cell Sci.94, 273-285.",
      "DOI": "10.1242/jcs.94.2.273"
    },
    {
      "key": "REF15",
      "doi-asserted-by": "crossref",
      "unstructured": "Hoops, H.J., and Witman, G.B. (1983). Outer doublet heterogeneity reveals structural polarity related to beat direction inChlamydomonasflagella.J. Cell Biol.97, 902-908.",
      "DOI": "10.1083/jcb.97.3.902"
    },
    {
      "key": "REF16",
      "doi-asserted-by": "crossref",
      "unstructured": "Huang, B., Ramanis, Z., Dutcher, S., and Luck, D.L. (1982). Uniflagellar mutants ofChlamydomonas: evidence for the role of basal bodies in transmission of positional information.Cell29, 745-753.",
      "DOI": "10.1016/0092-8674(82)90436-6"
    },
    {
      "key": "REF17",
      "doi-asserted-by": "crossref",
      "unstructured": "Johnson, U.G., and Porter, K.R. (1968). Fine structure of cell division inChlamydomonas reinhardtii.Basal bodies and microtubules.J. Cell Biol.38, 403-425.",
      "DOI": "10.1083/jcb.38.2.403"
    },
    {
      "key": "REF18",
      "doi-asserted-by": "crossref",
      "unstructured": "Kellogg, D.R., Moritz, M., and Alberts, B.A. (1994). The centrosome and cellular organization.Annu. Rev. Biochem.63, 639-674.",
      "DOI": "10.1146/annurev.bi.63.070194.003231"
    },
    {
      "key": "REF19",
      "doi-asserted-by": "crossref",
      "unstructured": "Kremer, J.R., Mastronarde, D.N., and McIntosh, J.R. (1996). Computer visualization of three-dimensional image data using IMOD.J. Struct. Biol.116, 71-76.",
      "DOI": "10.1006/jsbi.1996.0013"
    },
    {
      "key": "REF20",
      "doi-asserted-by": "crossref",
      "unstructured": "Ladinsky, M.S., Kremer, J.R., Furcinitti, P.S., McIntosh, J.R., and Howell, K.E. (1994). HVEM tomography of the trans-Golgi network: structural insights and identification of a lace-like vesicle coat.J. Cell Biol.127, 29-38.",
      "DOI": "10.1083/jcb.127.1.29"
    },
    {
      "key": "REF21",
      "doi-asserted-by": "crossref",
      "unstructured": "Ladinsky, M.S., Mastronarde, D.N., McIntosh, J.R., Howell, K.E., and Staehelin, L.A. (1999). Golgi structure in three dimensions: functional insights from the normal rat kidney cell.J. Cell Biol.144, 1135-1149.",
      "DOI": "10.1083/jcb.144.6.1135"
    },
    {
      "key": "REF22",
      "doi-asserted-by": "crossref",
      "unstructured": "Lange, B.M., and Gull, K. (1995). Molecular marker for centriole maturation in the mammalian cell cycle.J. Cell Biol.130, 919-927.",
      "DOI": "10.1083/jcb.130.4.919"
    },
    {
      "key": "REF23",
      "doi-asserted-by": "crossref",
      "unstructured": "Lechtreck, K-F., Teltenk\u00f6tter, A., and Grunow, A. (1999). A 210 kDa protein is located in a membrane-microtubule linker at the distal end of mature and nascent basal bodies.J. Cell Sci.112, 1633-1644.",
      "DOI": "10.1242/jcs.112.11.1633"
    },
    {
      "key": "REF24",
      "doi-asserted-by": "crossref",
      "unstructured": "LeDizet, M., and Piperno, G. (1986). Cytoplasmic microtubules containing acetylated alpha-tubulin inChlamydomonas reinhardtii: spatial arrangement and properties.J. Cell Biol.103, 13-22.",
      "DOI": "10.1083/jcb.103.1.13"
    },
    {
      "key": "REF25",
      "doi-asserted-by": "crossref",
      "unstructured": "Marsh, B.J., Mastronarde, D.N., Buttle, K.F., Howell, K.E., and McIntosh, J.R. (2001). Organellar relationships in the Golgi region of the pancreatic beta cell line, HIT-T15, visualized by high resolution electron tomography.Proc. Natl. Acad. Sci. USA98, 2399-2406.",
      "DOI": "10.1073/pnas.051631998"
    },
    {
      "key": "REF26",
      "doi-asserted-by": "crossref",
      "unstructured": "Mastronarde, D.N. (1997). Dual-axis tomography: an approach with alignment methods that preserve resolution.J. Struct. Biol.120, 343-352.",
      "DOI": "10.1006/jsbi.1997.3919"
    },
    {
      "key": "REF27",
      "doi-asserted-by": "crossref",
      "unstructured": "Moritz, M., Braunfeld, M.B., Fung, J.C., Sedat, J.W., Alberts, B.M., and Agard, D.A. (1995a). Three-dimensional structural characterization of centrosomes from earlyDrosophilaembryos.J. Cell Biol.130, 1149-1159.",
      "DOI": "10.1083/jcb.130.5.1149"
    },
    {
      "key": "REF28",
      "doi-asserted-by": "crossref",
      "unstructured": "Moritz, M., Braunfeld, M.B., Guenebaut, V., Heuser, J., and Agard, D.A. (2000). Structure of the \u03b3-tubulin ring complex: a template for microtubule nucleation.Nat. Cell Biol.2, 365-370.",
      "DOI": "10.1038/35014058"
    },
    {
      "key": "REF29",
      "doi-asserted-by": "crossref",
      "unstructured": "Moritz, M., Braunfeld, M.B., Sedat, J.W., Alberts, B., and Agard, D.A. (1995b). Microtuble nucleation by \u03b3-tubulin-containing rings in the centrosome.Nature378, 638-640.",
      "DOI": "10.1038/378638a0"
    },
    {
      "key": "atypb1",
      "doi-asserted-by": "crossref",
      "unstructured": "Oakley, B.R. (2000). An abundance of tubulins.Trends Cell. Biol.10, 537-542.",
      "DOI": "10.1016/S0962-8924(00)01857-2"
    },
    {
      "key": "REF31",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.10.6.2017"
    },
    {
      "key": "REF32",
      "doi-asserted-by": "crossref",
      "unstructured": "Piel, M., Meyer, P., Khodjakov, A., Reider, C.L., and Bornens, M. (2000). The respective contributions of mother and daughter centrioles to centrosome activity and behavior in vertebrate cells.J. Cell Biol.149, 317-329.",
      "DOI": "10.1083/jcb.149.2.317"
    },
    {
      "key": "REF33",
      "doi-asserted-by": "crossref",
      "unstructured": "Preble, A.M., Giddings, T.H., and Dutcher, S.K. (2000). Basal bodies and centrioles: their function and structure.Curr. Top. Dev. Biol.49, 207-233.",
      "DOI": "10.1016/S0070-2153(99)49010-6"
    },
    {
      "key": "REF34",
      "doi-asserted-by": "crossref",
      "unstructured": "Preble, A.M., Giddings, T.H., and Dutcher, S.K. (2001). Extragenic bypass supressors of mutations in the essential geneBLD2promote assembly of basal bodies with abnormal microtubules inChlamydomonas reinhardtii.Genetics157, 163-181.",
      "DOI": "10.1093/genetics/157.1.163"
    },
    {
      "key": "REF35",
      "doi-asserted-by": "crossref",
      "unstructured": "Quarmby, L.M., and Lohret, T.A. (1999). Microtubule severing.Cell Motil. Cytoskeleton43, 1-9.",
      "DOI": "10.1002/(SICI)1097-0169(1999)43:1<1::AID-CM1>3.0.CO;2-#"
    },
    {
      "key": "REF36",
      "doi-asserted-by": "crossref",
      "unstructured": "Ringo, D.L. (1967). Flagellar motion and fine structure of the flagellar apparatus inChlamydomonas.J. Cell Biol.33, 543-571.",
      "DOI": "10.1083/jcb.33.3.543"
    },
    {
      "key": "REF37",
      "doi-asserted-by": "crossref",
      "unstructured": "Salisbury, J.L., Baron, A.T., and Sanders, M.A. (1988). The centrinbased cytoskeleton ofChlamydomonas reinhardtii: distribution in interphase and mitotic cells.J. Cell Biol.107, 635-641.",
      "DOI": "10.1083/jcb.107.2.635"
    },
    {
      "key": "REF38",
      "doi-asserted-by": "crossref",
      "unstructured": "Sanders, M.A., and Salisbury, J.L. (1989). Centrin-mediated microtubule severing during flagellar excision inChlamydomonas reinhardtii.J. Cell Biol.108, 1751-1760.",
      "DOI": "10.1083/jcb.108.5.1751"
    },
    {
      "key": "REF39",
      "doi-asserted-by": "crossref",
      "unstructured": "Sanders, M.A., and Salisbury, J.L. (1994). Centrin plays an essential role in microtubule severing during flagellar excision inChlamydomonas reinhardtii.J. Cell Biol.124, 795-805.",
      "DOI": "10.1083/jcb.124.5.795"
    },
    {
      "key": "REF40",
      "doi-asserted-by": "crossref",
      "unstructured": "Schiebel E. (2000). Two new tubulins differ in a split decision.Nat. Cell Biol.2, E3-E4.",
      "DOI": "10.1038/71383"
    },
    {
      "key": "REF41",
      "doi-asserted-by": "crossref",
      "unstructured": "Silflow, C.D., Liu, B., LaVoie, M., Richardson, E.A., and Palevitz, B.A. (1999). \u03b3-Tubulin inChlamydomonas: characterization of the gene and localization of the gene product in cells.Cell Motil. Cytoskeleton42, 285-297.",
      "DOI": "10.1002/(SICI)1097-0169(1999)42:4<285::AID-CM3>3.0.CO;2-Z"
    },
    {
      "key": "REF42",
      "doi-asserted-by": "crossref",
      "unstructured": "Silflow, C. D., LaVoie, M., Tam L-W., Tousey, S., Sanders, M., Wu, W., Borodovsky, M., and Lefebvre, P.A. (2001). The Vfl1 protein inChlamydomonaslocalizes in a rotationally asymmetric pattern at the distal ends of basal bodies.J. Cell Biol.153, 63-74.",
      "DOI": "10.1083/jcb.153.1.63"
    },
    {
      "key": "atypb2",
      "doi-asserted-by": "crossref",
      "unstructured": "Smrzka, O.W., Delgehyr, N., and Bornens, M. (2000). Tissue-specific expression and subcellular localization of mammalian \u03b4-tubulin.Curr. Biol.10, 413-416.",
      "DOI": "10.1016/S0960-9822(00)00418-8"
    },
    {
      "key": "REF44",
      "doi-asserted-by": "crossref",
      "unstructured": "Taillon, B.E., Adler, S.A., Suhan, J.P., and Jarvik, J.W. (1992). Mutational analysis of centrin: an EF-hand protein associated with three distinct contractile fibers in the basal body apparatus ofChlamydomonas.J. Cell Biol.119, 1613-1624.",
      "DOI": "10.1083/jcb.119.6.1613"
    },
    {
      "key": "REF45",
      "doi-asserted-by": "crossref",
      "unstructured": "Wright, R.L., Chojnacki, B., and Jarvik, J.W. (1983). Abnormal basal-body number, location, and orientation in a striated fiber-defective mutant ofChlamydomonas reinhardtii.J. Cell Biol.96, 1697-1707.",
      "DOI": "10.1083/jcb.96.6.1697"
    },
    {
      "key": "REF46",
      "doi-asserted-by": "crossref",
      "unstructured": "Wright, R.L., Salisbury, J., and Jarvik, J.W. (1985). A nucleus-basal body connector inChlamydomonas reinhardtiithat may function in basal body localization or segregation.J. Cell Biol.101, 1903-1912.",
      "DOI": "10.1083/jcb.101.5.1903"
    }
  ],
  "container-title": "Molecular Biology of the Cell",
  "original-title": [],
  "language": "en",
  "deposited": {
    "date-parts": [
      [
        2021,
        6,
        6
      ]
    ],
    "date-time": "2021-06-06T05:45:21Z",
    "timestamp": 1622958321000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://www.molbiolcell.org/doi/10.1091/mbc.e02-11-0755"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2003,
        7
      ]
    ]
  },
  "references-count": 46,
  "journal-issue": {
    "issue": "7",
    "published-print": {
      "date-parts": [
        [
          2003,
          7
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1091/mbc.e02-11-0755"
  ],
  "URL": "http://dx.doi.org/10.1091/mbc.E02-11-0755",
  "relation": {},
  "ISSN": [
    "1059-1524",
    "1939-4586"
  ],
  "subject": [],
  "container-title-short": "MBoC",
  "published": {
    "date-parts": [
      [
        2003,
        7
      ]
    ]
  }
}