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First published online 8 November 2005
doi: 10.1242/jcs.02654

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Three distinct chromatin domains in telomere ends of polytene chromosomes in Drosophila melanogaster Tel mutants

Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, Novosibirsk, 630090, Russia

View larger version (136K): [in a new window]   Fig. 1. Telomere structure and distribution of proteins in polytene chromosomes X, 2R and 3R. (A) Electron microscopy. (B,E) FISH of TAS X probe. (C,F-I) Chromosome localization of chromatin proteins in telomeric regions: SUUR (C), PC (F), E(Z) (G), H3Me3K9 (H), H3Me3K27 (I). (D,E) Induction of swellings in UAS-SuUR+/+; Tel/ru h SuUR Sgs3-GAL4. (D) Orcein staining. (B,C,E-I) Merged image of the phase contrast (black and white) and immunofluorescence (red). Black arrowheads indicate the cap regions, white arrowheads show IH regions. Swellings are marked by arrows. Dashed lines, TAS repeats; solid lines, marker regions. Bars, 1 µm.

View larger version (77K): [in a new window]   Fig. 2. Telomere structure and protein localization in polytene chromosome arms 2L and 3L. (A) electron microscopy. (B,E) FISH of the 2L TAS probe. (C,F-K) Localization of antibodies to the chromatin proteins in telomeric regions: SUUR (C), PC (F,G), E(Z) (H,I), H3Me3K9 (J) and H3Me3K27 (K). (D and E) Induction of swellings in hybrids UAS-SuUR+/+; Tel/ru h SuUR Sgs3-GAL4; (D) orcein staining. Swellings are marked by arrows. Dashed lines, TAS repeats; solid lines, marker regions. Bars, 1 µm.

View larger version (96K): [in a new window]   Fig. 3. Localization of antibodies to the chromatin proteins in the telomeric regions of polytene chromosomes 2R (A-C,I,K,L), X (DH,J,M,O) and 3L (N). Capping proteins: (A) SUUR, (B) SU(VAR)3-7 and (C) HP2 localize to the chromosome tip. (D) H4AcK12 is not detected in telomeres. HeT-A/TAHRE/TART repeat recruits typical euchromatic proteins: (E) JIL-1, (F) Z4, (G) H3Me3K4 and Z4, both of which co-localize within this region. No productive transcription is detected within the HeT-A/TAHRE/TART region, since no signal for phospho-serine5 PolIIo is seen (H). Upon simultaneous immunodetection of cap- and HeT-A/TAHRE/TART-array-specific proteins, no overlap of the two signals is observed: HP2 and Z4 (I), HP1 and JIL-1 (J), HP1 and H3Me3K4 (K), HP1 and H3Me3K9 (L,M). (N) H3Me3K9 is detected within the HeT-A/TAHRE/TART region of Su(var)20503/Su(var)20505 larvae. (O) PC protein, recruited to the TAS repeats, and Z4 do not merge perfectly within the region of TAS repeats. Dashed lines, TAS repeats; solid lines, marker regions.

View larger version (39K): [in a new window]   Fig. 4. IdU incorporation dynamics in the telomere of Tel/+ polytene chromosome X. (Top) Hoechst staining of DNA; (Middle) IdU incorporation pattern; (Bottom) merged image. IdU incorporation at the discontinuous labeling phase: (A-C) all over the telomere; (D-F) the cap region still replicates; (G-I) telomeric regions no longer incorporate the label by the beginning of the late S phase; (J-L) replication of the chromosome tip is completed prior to the replication termination in the IH regions. Dashed lines, TAS repeats; solid lines, marker regions.

View larger version (150K): [in a new window]   Fig. 5. Telomeric associations in Tel are formed by the cap or tract of HeT-A/TAHRE/TART as revealed by electron microscopy. Bar, 1 µm.

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