Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Dec 1;90(23):11039–11043. doi: 10.1073/pnas.90.23.11039

Molecular cloning and centrosomal localization of human caltractin.

V D Lee 1, B Huang 1
PMCID: PMC47917  PMID: 8248209

Abstract

Caltractin, a 20-kDa calcium-binding protein, was previously purified and cloned at the DNA level from the unicellular green alga Chlamydomonas. It is a structural component of the basal body complex, the major microtubule-organizing center in Chlamydomonas and the functional homolog of the centrosome in the animal cell. Here we report the characterization of a cDNA encoding a human caltractin that shares a high degree of amino acid identity (70%) with its algal counterpart. Caltractin was identified in both HeLa and BHK cells as a 21-kDa polypeptide specifically localized to the centrosome of interphase and mitotic cells. The high level of conservation in the amino acid sequence of caltractin from algae to humans and its association with the major microtubule-organizing center in the cell suggest that caltractin plays a fundamental role in microtubule-organizing center structure and function.

Full text

PDF
11039

Images in this article

11041Fig. 2
on p.11041
11042Fig. 3
on p.11042
11042Fig. 4
on p.11042

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Asai D. J., Brokaw C. J., Thompson W. C., Wilson L. Two different monoclonal antibodies to alpha-tubulin inhibit the bending of reactivated sea urchin spermatozoa. Cell Motil. 1982;2(6):599–614. doi: 10.1002/cm.970020608. [DOI] [PubMed] [Google Scholar]
  2. Baron A. T., Greenwood T. M., Salisbury J. L. Localization of the centrin-related 165,000-Mr protein of PtK2 cells during the cell cycle. Cell Motil Cytoskeleton. 1991;18(1):1–14. doi: 10.1002/cm.970180102. [DOI] [PubMed] [Google Scholar]
  3. Baron A. T., Salisbury J. L. Identification and localization of a novel, cytoskeletal, centrosome-associated protein in PtK2 cells. J Cell Biol. 1988 Dec;107(6 Pt 2):2669–2678. doi: 10.1083/jcb.107.6.2669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baum P., Furlong C., Byers B. Yeast gene required for spindle pole body duplication: homology of its product with Ca2+-binding proteins. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5512–5516. doi: 10.1073/pnas.83.15.5512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ben-Chetrit E., Gandy B. J., Tan E. M., Sullivan K. F. Isolation and characterization of a cDNA clone encoding the 60-kD component of the human SS-A/Ro ribonucleoprotein autoantigen. J Clin Invest. 1989 Apr;83(4):1284–1292. doi: 10.1172/JCI114013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bolduc C., Lee V. D., Huang B. Beta-tubulin mutants of the unicellular green alga Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1988 Jan;85(1):131–135. doi: 10.1073/pnas.85.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brinkley B. R. Microtubule organizing centers. Annu Rev Cell Biol. 1985;1:145–172. doi: 10.1146/annurev.cb.01.110185.001045. [DOI] [PubMed] [Google Scholar]
  8. Gard D. L., Hafezi S., Zhang T., Doxsey S. J. Centrosome duplication continues in cycloheximide-treated Xenopus blastulae in the absence of a detectable cell cycle. J Cell Biol. 1990 Jun;110(6):2033–2042. doi: 10.1083/jcb.110.6.2033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gopalakrishna R., Anderson W. B. Ca2+-induced hydrophobic site on calmodulin: application for purification of calmodulin by phenyl-Sepharose affinity chromatography. Biochem Biophys Res Commun. 1982 Jan 29;104(2):830–836. doi: 10.1016/0006-291x(82)90712-4. [DOI] [PubMed] [Google Scholar]
  10. Huang B., Mengersen A., Lee V. D. Molecular cloning of cDNA for caltractin, a basal body-associated Ca2+-binding protein: homology in its protein sequence with calmodulin and the yeast CDC31 gene product. J Cell Biol. 1988 Jul;107(1):133–140. doi: 10.1083/jcb.107.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huang B., Watterson D. M., Lee V. D., Schibler M. J. Purification and characterization of a basal body-associated Ca2+-binding protein. J Cell Biol. 1988 Jul;107(1):121–131. doi: 10.1083/jcb.107.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lee V. D., Stapleton M., Huang B. Genomic structure of Chlamydomonas caltractin. Evidence for intron insertion suggests a probable genealogy for the EF-hand superfamily of proteins. J Mol Biol. 1991 Sep 5;221(1):175–191. [PubMed] [Google Scholar]
  14. Mazia D. The chromosome cycle and the centrosome cycle in the mitotic cycle. Int Rev Cytol. 1987;100:49–92. doi: 10.1016/s0074-7696(08)61698-8. [DOI] [PubMed] [Google Scholar]
  15. Moncrief N. D., Kretsinger R. H., Goodman M. Evolution of EF-hand calcium-modulated proteins. I. Relationships based on amino acid sequences. J Mol Evol. 1990 Jun;30(6):522–562. doi: 10.1007/BF02101108. [DOI] [PubMed] [Google Scholar]
  16. Moudjou M., Paintrand M., Vigues B., Bornens M. A human centrosomal protein is immunologically related to basal body-associated proteins from lower eucaryotes and is involved in the nucleation of microtubules. J Cell Biol. 1991 Oct;115(1):129–140. doi: 10.1083/jcb.115.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nakayama S., Moncrief N. D., Kretsinger R. H. Evolution of EF-hand calcium-modulated proteins. II. Domains of several subfamilies have diverse evolutionary histories. J Mol Evol. 1992 May;34(5):416–448. doi: 10.1007/BF00162998. [DOI] [PubMed] [Google Scholar]
  18. Paintrand M., Moudjou M., Delacroix H., Bornens M. Centrosome organization and centriole architecture: their sensitivity to divalent cations. J Struct Biol. 1992 Mar-Apr;108(2):107–128. doi: 10.1016/1047-8477(92)90011-x. [DOI] [PubMed] [Google Scholar]
  19. Salisbury J. L., Baron A. T., Coling D. E., Martindale V. E., Sanders M. A. Calcium-modulated contractile proteins associated with the eucaryotic centrosome. Cell Motil Cytoskeleton. 1986;6(2):193–197. doi: 10.1002/cm.970060218. [DOI] [PubMed] [Google Scholar]
  20. Salisbury J. L., Baron A. T., Sanders M. A. The centrin-based cytoskeleton of Chlamydomonas reinhardtii: distribution in interphase and mitotic cells. J Cell Biol. 1988 Aug;107(2):635–641. doi: 10.1083/jcb.107.2.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Salisbury J. L., Baron A., Surek B., Melkonian M. Striated flagellar roots: isolation and partial characterization of a calcium-modulated contractile organelle. J Cell Biol. 1984 Sep;99(3):962–970. doi: 10.1083/jcb.99.3.962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Salisbury J. L., Floyd G. L. Calcium-induced contraction of the rhizoplast of a quadriflagellate green alga. Science. 1978 Dec 1;202(4371):975–977. doi: 10.1126/science.202.4371.975. [DOI] [PubMed] [Google Scholar]
  23. Taillon B. E., Adler S. A., Suhan J. P., Jarvik J. W. Mutational analysis of centrin: an EF-hand protein associated with three distinct contractile fibers in the basal body apparatus of Chlamydomonas. J Cell Biol. 1992 Dec;119(6):1613–1624. doi: 10.1083/jcb.119.6.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tufty R. M., Kretsinger R. H. Troponin and parvalbumin calcium binding regions predicted in myosin light chain and T4 lysozyme. Science. 1975 Jan 17;187(4172):167–169. doi: 10.1126/science.1111094. [DOI] [PubMed] [Google Scholar]
  25. Wawrzynczak E. J., Perham R. N. Isolation and nucleotide sequence of a cDNA encoding human calmodulin. Biochem Int. 1984 Aug;9(2):177–185. [PubMed] [Google Scholar]
  26. Wright R. L., Salisbury J., Jarvik J. W. A nucleus-basal body connector in Chlamydomonas reinhardtii that may function in basal body localization or segregation. J Cell Biol. 1985 Nov;101(5 Pt 1):1903–1912. doi: 10.1083/jcb.101.5.1903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ye R. D., Wun T. C., Sadler J. E. cDNA cloning and expression in Escherichia coli of a plasminogen activator inhibitor from human placenta. J Biol Chem. 1987 Mar 15;262(8):3718–3725. [PubMed] [Google Scholar]
  28. Zhu J. K., Bressan R. A., Hasegawa P. M. An Atriplex nummularia cDNA with Sequence Relatedness to the Algal Caltractin Gene. Plant Physiol. 1992 Aug;99(4):1734–1735. doi: 10.1104/pp.99.4.1734. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES