Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes
In <i>Drosophila</i> salivary gland polytene chromosomes, a substantial portion of heterochromatin is underreplicated. The combination of mutations <i>SuUR<sup>ES</sup></i> and <i>Su(var)3-9<sup>06</sup></i> results in the polytenization of...
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2020-06-01
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| author | Tatyana D. Kolesnikova Alexandra V. Kolodyazhnaya Galina V. Pokholkova Veit Schubert Viktoria V. Dovgan Svetlana A. Romanenko Dmitry Yu. Prokopov Igor F. Zhimulev |
| author_facet | Tatyana D. Kolesnikova Alexandra V. Kolodyazhnaya Galina V. Pokholkova Veit Schubert Viktoria V. Dovgan Svetlana A. Romanenko Dmitry Yu. Prokopov Igor F. Zhimulev |
| author_sort | Tatyana D. Kolesnikova |
| collection | DOAJ |
| description | In <i>Drosophila</i> salivary gland polytene chromosomes, a substantial portion of heterochromatin is underreplicated. The combination of mutations <i>SuUR<sup>ES</sup></i> and <i>Su(var)3-9<sup>06</sup></i> results in the polytenization of a substantial fraction of unique and moderately repeated sequences but has almost no effect on satellite DNA replication. The Rap1 interacting factor 1 (Rif) protein is a conserved regulator of replication timing, and in <i>Drosophila</i>, it affects underreplication in polytene chromosomes. We compared the morphology of pericentromeric regions and labeling patterns of in situ hybridization of heterochromatin-specific DNA probes between wild-type salivary gland polytene chromosomes and the chromosomes of <i>Rif1</i> mutants and <i>SuUR Su(var)3-9<sup>06</sup></i> double mutants. We show that, despite general similarities, heterochromatin zones exist that are polytenized only in the <i>Rif1</i> mutants, and that there are zones that are under specific control of <i>Su(var)3-9</i>. In the <i>Rif1</i> mutants, we found additional polytenization of the largest blocks of satellite DNA (in particular, satellite 1.688 of chromosome X and simple satellites in chromosomes X and 4) as well as partial polytenization of chromosome Y. Data on pulsed incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into polytene chromosomes indicated that in the <i>Rif1</i> mutants, just as in the wild type, most of the heterochromatin becomes replicated during the late S phase. Nevertheless, a significantly increased number of heterochromatin replicons was noted. These results suggest that <i>Rif1</i> regulates the activation probability of heterochromatic origins in the satellite DNA region. |
| first_indexed | 2024-03-10T19:01:30Z |
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| language | English |
| last_indexed | 2024-03-10T19:01:30Z |
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| spelling | doaj.art-b29c9c0e26a24109a7dedff285dc94642023-11-20T04:24:51ZengMDPI AGCells2073-44092020-06-0196150110.3390/cells9061501Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene ChromosomesTatyana D. Kolesnikova0Alexandra V. Kolodyazhnaya1Galina V. Pokholkova2Veit Schubert3Viktoria V. Dovgan4Svetlana A. Romanenko5Dmitry Yu. Prokopov6Igor F. Zhimulev7Institute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaLeibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Seeland, GermanyInstitute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, 630090 Novosibirsk, RussiaIn <i>Drosophila</i> salivary gland polytene chromosomes, a substantial portion of heterochromatin is underreplicated. The combination of mutations <i>SuUR<sup>ES</sup></i> and <i>Su(var)3-9<sup>06</sup></i> results in the polytenization of a substantial fraction of unique and moderately repeated sequences but has almost no effect on satellite DNA replication. The Rap1 interacting factor 1 (Rif) protein is a conserved regulator of replication timing, and in <i>Drosophila</i>, it affects underreplication in polytene chromosomes. We compared the morphology of pericentromeric regions and labeling patterns of in situ hybridization of heterochromatin-specific DNA probes between wild-type salivary gland polytene chromosomes and the chromosomes of <i>Rif1</i> mutants and <i>SuUR Su(var)3-9<sup>06</sup></i> double mutants. We show that, despite general similarities, heterochromatin zones exist that are polytenized only in the <i>Rif1</i> mutants, and that there are zones that are under specific control of <i>Su(var)3-9</i>. In the <i>Rif1</i> mutants, we found additional polytenization of the largest blocks of satellite DNA (in particular, satellite 1.688 of chromosome X and simple satellites in chromosomes X and 4) as well as partial polytenization of chromosome Y. Data on pulsed incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into polytene chromosomes indicated that in the <i>Rif1</i> mutants, just as in the wild type, most of the heterochromatin becomes replicated during the late S phase. Nevertheless, a significantly increased number of heterochromatin replicons was noted. These results suggest that <i>Rif1</i> regulates the activation probability of heterochromatic origins in the satellite DNA region.https://www.mdpi.com/2073-4409/9/6/1501replication timing<i>Rif1</i><i>SuUR</i><i>Su(var)3-9</i><i>Drosophila melanogaster</i>polytene chromosomes |
| spellingShingle | Tatyana D. Kolesnikova Alexandra V. Kolodyazhnaya Galina V. Pokholkova Veit Schubert Viktoria V. Dovgan Svetlana A. Romanenko Dmitry Yu. Prokopov Igor F. Zhimulev Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes Cells replication timing <i>Rif1</i> <i>SuUR</i> <i>Su(var)3-9</i> <i>Drosophila melanogaster</i> polytene chromosomes |
| title | Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes |
| title_full | Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes |
| title_fullStr | Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes |
| title_full_unstemmed | Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes |
| title_short | Effects of Mutations in the <i>Drosophila melanogaster Rif1</i> Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes |
| title_sort | effects of mutations in the i drosophila melanogaster rif1 i gene on the replication and underreplication of pericentromeric heterochromatin in salivary gland polytene chromosomes |
| topic | replication timing <i>Rif1</i> <i>SuUR</i> <i>Su(var)3-9</i> <i>Drosophila melanogaster</i> polytene chromosomes |
| url | https://www.mdpi.com/2073-4409/9/6/1501 |
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