Cytoplasmic incompatibility in Drosophila: Biochemical and cellular analysis of factors influencing its expression
Chang, Wanlin
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Permalink
https://hdl.handle.net/2142/21121
Description
Title
Cytoplasmic incompatibility in Drosophila: Biochemical and cellular analysis of factors influencing its expression
Author(s)
Chang, Wanlin
Issue Date
1995
Doctoral Committee Chair(s)
Hager, Lowell P.
Department of Study
Biochemistry
Discipline
Biochemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Cell
Biology, Microbiology
Health Sciences, Immunology
Language
eng
Abstract
Wolbachia, an endocellular bacterium closely associated with cytoplasmic incompatibility in insects (CI), was purified from infected eggs and microinjected into two strains of D. melanogaster, a highly inbred laboratory strain (Oregon R), and a recently caught wild-type strain from Arizona. Infection status of the progeny from the microinjections was monitored by the polymerase chain reaction and indirect immunofluorescence using a Wolbachia specific monoclonal antibody. One infected line was established from the Arizona stock. This transinfected line expressed CI at significant, but slightly lower levels seen in the original host strain, D. simulans. The level of infection of Wolbachia in the transinfected D. melanogaster strain was estimated from bacterial counts in eggs from infected mothers and shown to be similar to those in the parental D. simulans host strain. These results suggest that (1) highly purified Wolbachia remain viable and capable of infecting a new host and (2) host factors may play an important role in the establishment and maintenance of the infected state and in the expression of cytoplasmic incompatibility.
In an effort to understand the biochemistry of CI, sperm from D. simulans-Riverside and tetracycline-treated D. simulans-Riverside (uninfected) were analyzed using gel electrophoresis. Results from SDS-PAGE, IEF, and 2-dimensional gel electrophoresis indicated that there exists an extra protein (not found in DSR sperm) that has a molecular weight of approximately 90 kD and a pI of approximately 4.5. Furthermore, sperm radiolabelling experiments suggest that this protein is phosphorylated. These results provided the first biochemical explanation of cytoplasmic incompatibility.
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