Temporal and spatial expression of the cell-cycle regulator cul-1 in Drosophila and its stimulation by

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V Filippov1, M Filippova, F Sehnal

  • 1Department of Cell Biology and Neuroscience Graduate Programs in Biochemistry and Molecular Biology, University of California, Riverside, CA 92521, USA.

The Journal of Experimental Biology
|August 23, 2000
PubMed

Insights

The Cul-1 ubiquitin ligase complex regulates cell division. The Drosophila Cul-1 gene can rescue yeast cell-cycle arrest, indicating conserved function despite differing mutation effects.

Area of Science:

  • Cell Biology
  • Genetics
  • Developmental Biology

Background:

  • The Cul-1 protein is a conserved component of ubiquitin ligase complexes.
  • These complexes target cell-cycle regulators for degradation, controlling cell proliferation.
  • Null mutations in the cul-1 gene exhibit distinct phenotypes in yeast (G1/S arrest) and nematodes (excessive cell division).

Purpose of the Study:

  • To isolate and characterize cul-1 orthologues in Drosophila melanogaster and mouse.
  • To investigate the functional conservation of the Drosophila cul-1 gene in yeast.
  • To analyze the expression patterns of the Drosophila cul-1 gene during development and in response to DNA damage.

Main Methods:

  • Isolation of cul-1 orthologues from Drosophila melanogaster and mouse.
  • Functional complementation assay using a yeast cul-1 mutant.
  • Analysis of Drosophila cul-1 gene expression at transcript and protein levels using RT-PCR and Western blotting.
  • Treatment of cultured cells with gamma-irradiation to assess transcriptional response.

Main Results:

  • The full-length Drosophila cul-1 gene successfully restored the G1/S-phase transition in the yeast mutant.
  • Drosophila cul-1 is maternally supplied as protein but not RNA; zygotic transcription begins early in embryogenesis.
  • Cul-1 transcription increased in cultured cells exposed to lethal gamma-irradiation.

Conclusions:

  • The Drosophila cul-1 gene functionally complements yeast mutations, demonstrating conserved roles in cell-cycle regulation.
  • The developmental expression pattern suggests maternal contribution and subsequent zygotic regulation of Cul-1.
  • Cul-1 expression is responsive to DNA damage, hinting at its involvement in DNA repair pathways.

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