Identification and Characterization of Proteins Involved in Nuclear Organization Using Drosophila GFP Protein Trap Lines

Margaret Rohrbaugh, Alyssia Clore, Julia Davis, Sharonta Johnson, Brian Jones, Keith Jones, Joanne Kim, Bramwel Kithuka, Krystal Lunsford, Joy Mitchell, Brian Mott, Edward Ramos, Maza R. Tchedou, Gilbert Acosta, Mark Araujo, Stuart Cushing, Gabriel Duffy, Felicia Graves, Kyler Griffin, B. V. GurudattaDeaundra Jackson, Denis Jaimes, Kendall Jamison, Khali Jones, Dhaujee Kelley, Marquita Kilgore, Derica Laramore, Thuy Le, Bakhtawar Mazhar, Muhammad M. Mazhar, Britney McCrary, Teanndras Miller, Celethia Moreland, Alex Mullins, Elyas Munye, Sheila Okoorie, Elisha Pittman, Nikkita Roberts, De'Warren Rose, Alex Rowland, Anwar Shagarabi, Jamela Smith, Tayler Stallworth, Nicole Stroud, Elizabeth Sung, Kai Sung, Naomi Takenaka, Eduardo Torre, Jarvis Veira, Kim Vu

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Background: Strains from a collection of Drosophila GFP protein trap lines express GFP in the normal tissues where the endogenous protein is present. This collection can be used to screen for proteins distributed in the nucleus in a non-uniform pattern. Methodology/Principal Findings: We analyzed four lines that show peripheral or punctate nuclear staining. One of these lines affects an uncharacterized gene named CG11138. The CG11138 protein shows a punctate distribution in the nuclear periphery similar to that of Drosophila insulator proteins but does not co-localize with known insulators. Interestingly, mutations in Lamin proteins result in alterations in CG11138 localization, suggesting that this protein may be a novel component of the nuclear lamina. A second line affects the Decondensation factor 31 (Df31) gene, which encodes a protein with a unique nuclear distribution that appears to segment the nucleus into four different compartments. The X-chromosome of males is confined to one of these compartments. We also find that Drosophila Nucleoplasmin (dNlp) is present in regions of active transcription. Heat shock leads to loss of dNlp from previously transcribed regions of polytene chromosome without redistribution to the heat shock genes. Analysis of Stonewall (Stwl), a protein previously found to be necessary for the maintenance of germline stem cells, shows that Stwl is present in a punctate pattern in the nucleus that partially overlaps with that of known insulator proteins. Finally we show that Stwl, dNlp, and Df31 form part of a highly interactive network. The properties of other components of this network may help understand the role of these proteins in nuclear biology. Conclusions/Significance: These results establish screening of GFP protein trap alleles as a strategy to identify factors with novel cellular functions. Information gained from the analysis of CG11138 Stwl, dNlp, and Df31 sets the stage for future studies of these proteins.

Original languageEnglish (US)
Article numbere53091
JournalPloS one
Volume8
Issue number1
DOIs
StatePublished - Jan 16 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Rohrbaugh, M., Clore, A., Davis, J., Johnson, S., Jones, B., Jones, K., Kim, J., Kithuka, B., Lunsford, K., Mitchell, J., Mott, B., Ramos, E., Tchedou, M. R., Acosta, G., Araujo, M., Cushing, S., Duffy, G., Graves, F., Griffin, K., ... Vu, K. (2013). Identification and Characterization of Proteins Involved in Nuclear Organization Using Drosophila GFP Protein Trap Lines. PloS one, 8(1), [e53091]. https://doi.org/10.1371/journal.pone.0053091