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. 2014;8(2):108-12.
doi: 10.4161/fly.28312.

In vivo analysis of a fluorescent SUMO fusion in transgenic Drosophila

Marion Bocksberger  1 , François Karch, Jean-Michel Gibert
Affiliations

In vivo analysis of a fluorescent SUMO fusion in transgenic Drosophila

Marion Bocksberger et al. Fly (Austin). 2014.

Abstract

Sumoylation, the covalent attachment of SUMO, a 90 amino acid peptide related to ubiquitin, is a major modulator of protein functions. Fluorescent SUMO protein fusions have been used in cell cultures to visualize SUMO in vivo but not in multicellular organisms. We generated a transgenic line of Drosophila expressing an mCherry-SUMO fusion. We analyzed its pattern in vivo in salivary gland nuclei expressing Venus-HP1 to recognize the different chromatin components (Chromocenter, chromosome IV). We compared it to SUMO immunostaining on squashed polytene chromosomes and observed similar patterns. In addition to the previously reported SUMO localizations (chromosome arms and chromocenter), we identify 2 intense binding sites: the fourth chromosome telomere and the DAPI-bright band in the region 81F.

Keywords: Drosophila; SUMO; chromatin; chromosome; fluorescent fusion.

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Figures

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Figure 1. In vivo observation of mCherry-SUMO in salivary glands (A and B). BB: a bright band is observed in most nuclei. (C–E) In vivo observation of mCherry-SUMO (CandE; red) and Venus-HP1 (D and E; green) in a salivary gland nucleus. (E) Merged. BB: mCherry-SUMO bright band. TIV: SUMO staining on chromosome IV telomere. Chr: chromocenter. IV: chromosome IV.
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Figure 2. Immunostaining for SUMO (A, C, D, and F; red) on squashed salivary gland wild-type polytene chromosomes. DNA is stained with DAPI (blue). (C and F) Merged. DB: DAPI bright band. Chr: chromocenter. TIV: chromosome IV telomere. IV: chromosome IV. The SUMO signal was adjusted not to saturate on the DAPI bright band, the telomere of the fourth chromosome and the chromocenter, so it appears weak on chromosome arms.
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