Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 461K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Karakesisoglou, I. Articles by Schleicher, M. Search for Related Content PubMed PubMed Citation Articles by Karakesisoglou, I. Articles by Schleicher, M. Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/1999//167 $5.00
The Journal of Cell Biology, Volume 145, Number 1, , 1999 167-181

Identification of a Suppressor of the Dictyostelium Profilin-minus Phenotype as a CD36/LIMP-II Homologue

Iakowos Karakesisoglou^*, Klaus-Peter Janssen^*, Ludwig Eichinger^*, Angelika A. Noegel, and Michael Schleicher^*

^* A.-Butenandt-Institut für Zellbiologie, Ludwig-Maximilians-Universität, 80336 München, Germany; and Institut für Biochemie I, Universität zu Köln, 50931 Köln, Germany

Profilin is an ubiquitous G-actin binding protein in eukaryotic cells. Lack of both profilin isoforms in Dictyostelium discoideum resulted in impaired cytokinesis and an arrest in development. A restriction enzyme–mediated integration approach was applied to profilin-minus cells to identify suppressor mutants for the developmental phenotype. A mutant with wild-type–like development and restored cytokinesis was isolated. The gene affected was found to code for an integral membrane glycoprotein of a predicted size of 88 kD containing two transmembrane domains, one at the NH₂ terminus and the other at the COOH terminus. It is homologous to mammalian CD36/LIMP-II and represents the first member of this family in D. discoideum, therefore the name DdLIMP is proposed. Targeted disruption of the lmpA gene in the profilin-minus background also rescued the mutant phenotype. Immunofluorescence revealed a localization in vesicles and ringlike structures on the cell surface. Partially purified DdLIMP bound specifically to PIP₂ in sedimentation and gel filtration assays. A direct interaction between DdLIMP and profilin could not be detected, and it is unclear how far upstream in a regulatory cascade DdLIMP might be positioned. However, the PIP₂ binding of DdLIMP points towards a function via the phosphatidylinositol pathway, a major regulator of profilin.

Key Words: cytoskeleton • LIMP-II • CD36 • profilin-suppressor • phosphatidylinositides

Abbreviations used in this paper: DAPI, 4,6-diamidino-2-phenylindole; ICAM, intercellular cell adhesion molecules; LIMP, lysosomal integral membrane protein; PC, phosphatidylcholine; PI, phosphatidylinositol; PS, phosphatidylserine; RB, REMI BamHI mutant; REMI, restriction enzyme–mediated integration.

I. Karakesisoglou's present address is Howard Hughes Medical Institute, Department of Molecular Genetics, Cell Biology, Biochemistry, and Molecular Biology, The University of Chicago, Chicago, IL 60637.

The work was supported by grants from the Deutsche Forschungsgemeinschaft to A.A. Noegel and M. Schleicher, and by funds from the Fonds der Chemischen Industrie and the Friedrich-Baur-Stiftung.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Facebook

Reddit

Technorati

Twitter

This article has been cited by other articles:

• Gareus, R., Di Nardo, A., Rybin, V., Witke, W. (2006). Mouse Profilin 2 Regulates Endocytosis and Competes with SH3 Ligand Binding to Dynamin 1. J. Biol. Chem. 281: 2803-2811 [Abstract] [Full Text]  
• Gamp, A.-C., Tanaka, Y., Lullmann-Rauch, R., Wittke, D., D'Hooge, R., De Deyn, P. P., Moser, T., Maier, H., Hartmann, D., Reiss, K., Illert, A.-L., von Figura, K., Saftig, P. (2003). LIMP-2/LGP85 deficiency causes ureteric pelvic junction obstruction, deafness and peripheral neuropathy in mice. Hum Mol Genet 12: 631-646 [Abstract] [Full Text]  
• Gotthardt, D., Warnatz, H. J., Henschel, O., Bruckert, F., Schleicher, M., Soldati, T. (2002). High-Resolution Dissection of Phagosome Maturation Reveals Distinct Membrane Trafficking Phases. Mol. Biol. Cell 13: 3508-3520 [Abstract] [Full Text]  
• Harris, E., Wang, N., Wu, W.-l, Weatherford, A., De Lozanne, A., Cardelli, J. (2002). Dictyostelium LvsB Mutants Model the Lysosomal Defects Associated with Chediak-Higashi Syndrome. Mol. Biol. Cell 13: 656-669 [Abstract] [Full Text]  
• Janssen, K.-P., Rost, R., Eichinger, L., Schleicher, M. (2001). Characterization of CD36/LIMPII Homologues in Dictyostelium discoideum. J. Biol. Chem. 276: 38899-38910 [Abstract] [Full Text]  
• Rupper, A., Lee, K., Knecht, D., Cardelli, J. (2001). Sequential Activities of Phosphoinositide 3-Kinase, PKB/Akt, and Rab7 during Macropinosome Formation in Dictyostelium. Mol. Biol. Cell 12: 2813-2824 [Abstract] [Full Text]  
• McKinney, E. C., Kandasamy, M. K., Meagher, R. B. (2001). Small Changes in the Regulation of One Arabidopsis Profilin Isovariant, PRF1, Alter Seedling Development. Plant Cell 13: 1179-1191 [Abstract] [Full Text]  
• Rupper, A., Grove, B., Cardelli, J. (2001). Rab7 regulates phagosome maturation in Dictyostelium. J. Cell Sci. 114: 2449-2460 [Abstract] [Full Text]  
• Rupper, A., Rodriguez-Paris, J., Grove, B., Cardelli, J. (2001). p110-related PI 3-kinases regulate phagosome-phagosome fusion and phagosomal pH through a PKB/Akt dependent pathway in Dictyostelium. J. Cell Sci. 114: 1283-1295 [Abstract]  
• Temesvari, L., Zhang, L., Fodera, B., Janssen, K.-P., Schleicher, M., Cardelli, J. A. (2000). Inactivation of lmpA, Encoding a LIMPII-related Endosomal Protein, Suppresses the Internalization and Endosomal Trafficking Defects in Profilin-null Mutants. Mol. Biol. Cell 11: 2019-2031 [Abstract] [Full Text]  
• Noegel, A., Schleicher, M (2000). The actin cytoskeleton of Dictyostelium: a story told by mutants. J. Cell Sci. 113: 759-766 [Abstract]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents