Andrea Pereira, Ph.D.

Academic Role: Research Assistant Professor

Faculty Appointment(s) In:
   Molecular Genetics and Microbiology

Other Affiliation(s):
   Cell Biology

Microtubule motor proteins of the kinesin superfamily

The microtubule motor proteins of the kinesin superfamily are known to be responsible for numerous kinds of motile events that occur during cell division. The main research goal of my lab is to extend the understanding of spindle assembly and function through the genetic analysis of several of these motor proteins: the Drosophila kinesin-like protein 67A (KLP67A and human homolog, kif18), KLP61F (human homolog hsEG5) as well as a novel cytoskeletal protein BSG25D.

Immunolocalization data of KLP67A reveals an interaction with both mitochondria and Wolbachia bacteria that are bound to the astral fibers of the spindle. This interaction is necessary for proper spindle assembly and function during cell division as demonstrated by a KLP67A mutant phenotype analysis. Current work on KLP67A is focused on understanding the molecular details of the motor-spindle-mitochondria complex. The second KLP that we are interested in is KLP61F. Although we have previously demonstrated that KLP61F has a role in spindle assembly, it is unclear as to whether this motor is initiating pole separation, stabilizing the spindle once formed and/or also powering spindle fiber sliding during anaphase movements. To address these questions, we have created a temperature sensitive allele of KLP61F. Our plan now is to observe the effects of this mutation on spindle assembly and chromosome movements. Mutations in KLP61F as well as KLP67A are also being used to illuminate the role of these molecular motors in the highly specialized female meiotic spindle.

In fact we have new and exciting data that points to a role of KLP67A in female meiotic spindle assembly. Together, these studies will increase our knowledge of cell division leading to better understanding of the steps that go awry during normal cellular development as well as in cancerous cells.

Figures

Model of astral forces. The distribution of KLPs, KLP61F and KLP67A on astral and spindle microtubules is indicated. The effects of mutations in KLP61F and KLP67A in spindle assembly are shown. Mutations in KLP61F prevent centrosome separation in larval neuroblasts The abnormal length and shape of the KLP67A mutant spindle is speculated to be due to improper centrosome migration during prometaphase.

Kinesin like proteins are involved in various aspects of cell division and show homology to the motor domain of conventional kinesin, an ATPase motor used to move organelles on microtubule tracks. We are working on a kinesin like protein, KLP67A in Drosophila melanogaster. KLP67A localizes to the ends of astral microtubules where it is associated with mitochondria and an endosymbiont, Wolbachia. Wolbachia is a maternally inherited endosymbiotic bacteria that manipulates host reproduction to increase the number of infected individuals within the population. Various mutants have been made and studied to understand the function of KLP67A. Also, yeast two hybrid and coimmunoprecipitation procedures are used to study the proteins interacting with KLP67A.

The figure shows the cell cycle expression of KLP67A in embryos - Early blastoderm embryos were stained with KLP67A (red signal) and a-tubulin antibodies (green signal). DNA is shown in blue. A, Prophase B, Metaphase C, Telophase.

Immunolocalization of KLP67A in CHO cells (vertebrate kidney cells). Cells were labelled with anti-KLP67A antibodies (a) and mitochondria specific dye Mitotracker (b). The pattern in both the cases is similar suggesting that KLP67A co localize with mitochondria. This indicate that KLP67A might be interacting with mitochondria.

Recent Publications

Gargazino, V, Pereira, A.J., Laurenti, P, Graba, Y, Levis, R, LeParco, Y, and Pradel, J (1992). Cell-lineage specific expression of modulo, a dose-dependent modifier of variegation in Drosophila. EMBO J. 11: 4471-4479

Pereira, A.J., Doshen, J, Tanaka, E, and Goldstein, LSB (1992). Genetic Analysis of a Drosophila microtubule-associated protein. J. Cell Biol 116: 377-383

Heck, MMS, Pereira, A.J., Pesavento, P, Yannoni, Y, Spradling, AC, and Goldstein, LSB (1993). The kinesin-like protein KLP61F is essential for mitosis in Drosophila. J. Cell Biol. 123: 665-679

Pereira, A.J., and Goldstein, L.S.B. (1993). Drosophila 205K MAP. In Guidebook to the Cytoskeletal, Extracellular Matrix and Adhesion Proteins. Oxford University Press, T Kreis and R Vale, Editors.

Pereira, A.J., and Goldstein, LSB (1994). The Kinesin Superfamily. In: JS Hyams and CW Lloyd (ed.) Microtubules. Wiley-Liss, pp. 269-284

Dalby, B, Pereira, A.J., and Goldstein, LSB (1995). An inverse PCR Screen for the detection of P Element insertions in cloned genomic intervals in Drosophila melanogaster. Genetics 139: 757-766

Barton, NR, Pereira, A.J., and Goldstein, LSB (1995). Motor activity and mitotic spindle localization of the Drosophila kinesin-like protein KLP61F. Mol. Biol. Cell 6: 1563-1574

Pereira, A.J., Dalby, B, Stewart, RJ, Doxsey, S D and Goldstein, LSB (1997). Mitochondrial association of a plus-end directed microtubule motor expressed during mitosis in Drosophila. J. Cell Biol. 136: 1081-1090

Gandhi, R., Bonaccorsi, S. Wentworth, Gatti, M, and Pereira, A. J. (2002) Interaction of a novel cytoskeletal protein with the kinesin-like protein, KLP67A (in preparation)

Gandhi, R., Bonaccorsi, S. Wentworth, Doxsey S., Gatti, M, and Pereira, A. J. (2002) Genetic evidence for the requirement of a mitochondria associated motor in spindle assembly. (manuscript in preparation)

Potential Rotation Projects

• Ultrastructure studies of the mitochondria-Wolbachia interaction
• RNAi inhibition studies of the human homologs using cultured cells
• Genetic characterization of a KLP67A interacting protein.
• Genetic modifier screen for klp67a interacting proteins.

Laboratory Staff

Academic Background

Ph.D, 1987 Brandeis University, Waltham MA

Postdoctoral fellow - Harvard University

Office: 770B1
Phone: 508-856-6236
E-mail: Andrea.Pereira@umassmed.edu

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