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First published online 24 February 2009
doi: 10.1242/jcs.035097

Journal of Cell Science 122, 769-774 (2009)
Published by The Company of Biologists 2009
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A product of the bicistronic Drosophila melanogaster gene CG31241, which also encodes a trimethylguanosine synthase, plays a role in telomere protection

Orban Komonyi1, Tamas Schauer1, Gabor Papai2, Peter Deak2 and Imre M. Boros1,2,*

1 Chromatin Research Group of HAS, Department of Biochemistry and Molecular Biology, University of Szeged, Közép fasor 52, H-6726 Szeged, Hungary
2 Institute of Biochemistry, Biological Research Center, Temesvari krt.62, H-6726 Szeged, Hungary


Figure 1
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  		Fig. 1. (A) Structure of the CG31241 gene, its alleles and transgenes with the associated phenotypes. Shaded boxes indicate the two ORFs of CG31241 encoding DTL and TGS1. The gap in uORF(DTL) box indicates the position of an intron. P(967) is a P-element insertion in the 5'-untranslated region of CG31241. d192, d58 and d189 are null alleles for both DTL and TGS1 functions. Breaks enclosed by parentheses indicate the positions and extensions of deletions. Casper-DTL, Casper-DTLum and Casper-DTLdm are transgenes that carry wild-type CG31241, a nonsense mutation in the DTL coding region and a deletion in the TGS1-coding region, respectively. pUAST-DTL and pUAST-TGS1 are cDNAs corresponding to the CG31241 uORF and dORF, respectively. (B) Rough eye (2), missing thoracic bristles (4, arrow), abnormal development of veins (6, arrowheads), scalloped wings (7, arrowhead) are characteristic for animals carrying the Casper-DTLum transgene and the hypomorph P(967) CG31241 allele. (1, 3 and 5 show corresponding controls). (C) In CG31241-null larvae, the size of imaginal discs (1, eye-antenna; 2, wing disc) is severely reduced compared with similar age controls (left, wild type; right, mutant). (D) Acridine orange staining of the imaginal discs indicates increased apoptosis in CG31241-null mutants. 1, wild type; 2, mutant wing disc. (E) Orcein-stained mitotic spreads of CG31241 neuroblasts display telomere associations. Loss of both DTL and TGS1 functions in d192/d189 cells (a-d), or DTL function in P[DTLum]/+; d192/d189 cells (g-j), but not the loss of TGS1 function alone in P[DTLdm]/+; d192/d189 cells (e,f) causes defective mitotic chromosomes and frequent telomere associations.

 

Figure 2
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  		Fig. 2. HOAP and HP1 binding to dtl chromosomes and frequency of different telomere associations in dtl neuroblasts. (A) HOAP-specific staining of CG31241 (P[DTLum]/+; d58/d189) mitotic chromosomes in larval neuroblasts. Chromosome end attachment types are indicated by arrows and numbers, indicating: intersister (1), single non-sister (2), double (3) and other (4) attachments. (B) Average frequencies for different types of telomere associations (TA) in dtl-null mutants (n>200). (C) Wild-type (left) and CG31241 (P[DTLum]/+; d58/d189) (right) polytene chromosomes immunostained with HOAP-specific antibody (red) and DAPI (blue). (D) HP-1-specific staining of wild-type (left) and CG31241 (P[DTLum]/+; d58/d189) (right) polytene chromosomes.

 

Figure 3
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  		Fig. 3. Stained polytene chromosomes of wild-type L3 larvae ectopically expressing DTL-Flag. On each panel merged DAPI (blue) and Flag-specific antibody stained (red) images are shown. (A) Chromosome spread showing localization of DTL-Flag signals at specific interbands. (B) Enlarged 2L chromosome ends of wild-type (top) and DTL-Flag-expressing animals (bottom). (C) Chromosome ends of DTL-Flag-expressing animals.

 

Figure 4
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  		Fig. 4. Genetic interaction between tefu/atm and mei-41/atr and CG31241 alleles. (A) Lethality phases of single and double mutants. (B,C) Frequency of anaphase bridges in tefu/atm and CG31241 single and double mutant neuroblasts. Error bars indicate range; 150 or more anaphases were scored for each genotype, each obtained from three independent crosses. P(967) is a hypomorph, d192 is a null allele of CG31241. Mei-41/atr mutations suppress CG31241 phenotype with respect to both developmental arrest (D), and number of anaphase bridges (E,F). d58 is null allele of CG31241, mei-41D9 and mei-41RT1 are weak and strong alleles of mei-41/atr, respectively. A detailed description of the crosses performed and the genotypes analyzed is given in Materials and Methods.

 

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© The Company of Biologists Ltd 2009