Evidence of the Physical Interaction between Rpl22 and the Transposable Element Doc5, a Heterochromatic Transposon of Drosophila melanogaster
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plasmids Construction
2.2. PCR Amplification
2.3. One Hybrid Screening
2.4. Protein Expression and Purification
2.5. Electrophoretic Mobility Shift Assay (EMSA)
2.6. Fluorescence In Situ Hybridization and Immunofluorescence on Polytene Chromosomes
2.7. Other Methods
3. Results
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer | Sequence | Usage |
---|---|---|
ADread | 5′-CTATTCGATGATGAAGAT-3′ | sequencing |
pACT2seq | 5′-TACCACTACAATGGATG-3′ | sequencing |
pACT2 up | 5′-CTATTCGATGATGAAGATACCCCACCAAACCC-3′ | Amplification/cloning |
pACT2 low | 5′-GTGAACTTGCGGGGTTTTTCAGTATCTACGAT-3′ | Amplification/cloning |
His1_up | 5′-GAGGCCCTTTCGTCTTCAA-3′ | Amplification/cloning |
His1_low | 5′-CTAGGGCTTTCTGCTCTGTCATCT-3′ | Amplification/cloning |
Doc5_up | 5′-ACGGCTATTATTGTTTCTTATTGCT-3′ | Amplification/cloning |
Doc5_low | 5′-TTATCCTCATCCCTTATCCTATGT-3′ | Amplification/cloning |
pETup | 5′-CACCATGGCTTACCCATA-3′ | Amplification/cloning |
pETlow | 5′-ATAAAAGAAGGCAAAACGATG-3′ | Amplification/cloning |
H5low | 5′-CTAACGCAGCACGTTCTTCTT-3′ | Amplification/cloning |
L22up | 5′-CACCAAGGTGGTCAAGAAGAA-3′ | Amplification/cloning |
Subject Accession | Start | End | Chromosome | % Identity | Alignment Length | Evalue | Bit Score | Chromosome Map Position |
---|---|---|---|---|---|---|---|---|
NT_033779.5 | 23,430,152 | 23,429,509 | 2L | 85.891 | 645 | 0 | 643 | h35–36 |
NT_033779.5 | 23,037,532 | 23,037,717 | 2L | 91.237 | 194 | 2.03 × 10−67 | 257 | h35–36 |
NW_001845128.1 | 3990 | 4435 | 2CEN | 100 | 446 | 0 | 824 | deep het |
NW_001845128.1 | 3875 | 3990 | 2CEN | 100 | 116 | 1.24 × 10−54 | 215 | deep het |
NW_001844967.1 | 11,017 | 10,572 | 2CEN | 100 | 446 | 0 | 824 | deep het |
NW_001844967.1 | 11,132 | 11,017 | 2CEN | 100 | 116 | 1.24 × 10−54 | 215 | deep het |
NW_007931075.1 | 7023 | 6491 | 2CEN | 82.655 | 565 | 2.43 × 10−121 | 436 | deep het |
NW_007931075.1 | 9914 | 9382 | 2CEN | 82.655 | 565 | 2.43 × 10−121 | 436 | deep het |
NT_033778.4 | 396,636 | 397,234 | 2R | 99.332 | 599 | 0 | 1083 | h41–h44 |
NT_033778.4 | 165,422 | 164,833 | 2R | 95.326 | 599 | 0 | 942 | h41–h44 |
NT_033778.4 | 74,075 | 74,637 | 2R | 92.833 | 586 | 0 | 824 | h41–h44 |
NT_033778.4 | 298,134 | 297,689 | 2R | 100 | 446 | 0 | 824 | h41–h44 |
NT_033778.4 | 872,342 | 871,810 | 2R | 82.655 | 565 | 2.43 × 10−121 | 436 | h41–h44 |
NT_033778.4 | 875,233 | 874,701 | 2R | 82.655 | 565 | 2.43 × 10−121 | 436 | h41–h44 |
NT_033778.4 | 1,413,093 | 1,413,384 | 2R | 91.333 | 300 | 2.47 × 10−111 | 403 | h45 |
NT_033778.4 | 3,518,328 | 3,518,018 | 2R | 88.179 | 313 | 1.95 × 10−97 | 357 | h41–h44 |
NT_033778.4 | 5,012,337 | 5,012,064 | 2R | 82.818 | 291 | 4.43 × 10−59 | 230 | h46 |
NT_033778.4 | 298,249 | 298,134 | 2R | 100 | 116 | 1.24 × 10−54 | 215 | h41-h44 |
NT_033778.4 | 5,012,596 | 5,012,448 | 2R | 84.302 | 172 | 4.59 × 10−34 | 147 | h46 |
NT_037436.4 | 24,877,579 | 24,878,162 | 3L | 87.081 | 596 | 0.0 | 656 | h49 |
NT_037436.4 | 24,914,416 | 24,913,834 | 3L | 86.745 | 596 | 0.0 | 645 | h49 |
NT_037436.4 | 24,944,076 | 24,944,602 | 3L | 87.199 | 539 | 2.93 × 10−170 | 599 | h49 |
NT_037436.4 | 24,461,859 | 24,462,331 | 3L | 85.443 | 474 | 6.71 × 10−127 | 455 | h47 |
NT_037436.4 | 23,664,082 | 23,663,846 | 3L | 94.583 | 240 | 9.00 × 10−101 | 368 | 80F9 |
NT_037436.4 | 23,663,844 | 23,663,639 | 3L | 92.754 | 207 | 1.20 × 10−79 | 298 | 80F9 |
NT_037436.4 | 25,490,094 | 25,490,271 | 3L | 94.382 | 178 | 2.02 × 10−72 | 274 | h49–h50 |
NT_037436.4 | 27,913,043 | 27,913,159 | 3L | 93.277 | 119 | 7.57 × 10−42 | 172 | h51 |
NT_037436.4 | 24,502,780 | 24,502,891 | 3L | 92.035 | 113 | 2.12 × 10−37 | 158 | h48 |
NT_037436.4 | 27,912,886 | 27,913,038 | 3L | 81.609 | 174 | 2.15 × 10−27 | 124 | h51 |
NT_033777.3 | 646,928 | 646,337 | 3R | 86.612 | 605 | 0.0 | 649 | h54–h56 |
NT_033777.3 | 4,042,323 | 4,042,066 | 3R | 94.961 | 258 | 6.86 × 10−112 | 405 | 81F |
NT_033777.3 | 1,401,453 | 1,401,023 | 3R | 83.991 | 431 | 1.50 × 10−103 | 377 | h54–h56 |
NT_033777.3 | 4,050,664 | 4,050,455 | 3R | 95.238 | 210 | 3.28 × 10−90 | 333 | 81F |
NT_033777.3 | 3,992,575 | 3,992,365 | 3R | 94.787 | 211 | 1.53 × 10−88 | 327 | 81F |
NT_033777.3 | 4,039,954 | 4,039,769 | 3R | 91.710 | 193 | 1.57 × 10−68 | 261 | 81F |
NT_033777.3 | 2,453,123 | 2,453,400 | 3R | 80.357 | 280 | 4.53 × 10−44 | 180 | h56 |
NT_033777.3 | 2,453,399 | 2,453,508 | 3R | 92.793 | 111 | 5.90 × 10−38 | 159 | h56 |
NW_001845051.1 | 2554 | 2831 | UNK | 80.357 | 280 | 4.53 × 10−44 | 180 | deep het |
NW_001845051.1 | 2830 | 2934 | UNK | 89.189 | 111 | 1.29 × 10−29 | 132 | deep het |
Gene Name | FlyBase ID | Function | Inferred by | Reference |
---|---|---|---|---|
vig | FBgn0024183 | Heterochromatin organization | Co-IP | [42] |
AGO1 | FBgn0262739 | transcriptional repression | Co-IP | [42] |
AGO2 | FBgn0087035 | transcriptional repression | Co-IP | [42] |
vig2 | FBgn0046214 | Heterochromatin organization | Mass-spec | [43] |
Fmr1 | FBgn0028734 | piRNA biogenesis | Co-IP | [42] |
ban | FBgn0262451 | piRNA biogenesis | Co-IP | [42] |
esi2 | FBgn0285992 | Unknown | Co-IP | [42] |
smt3 | FBgn0264922 | mitosis | Co-IP | [31] |
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Berloco, M.F.; Minervini, C.F.; Moschetti, R.; Palazzo, A.; Viggiano, L.; Marsano, R.M. Evidence of the Physical Interaction between Rpl22 and the Transposable Element Doc5, a Heterochromatic Transposon of Drosophila melanogaster. Genes 2021, 12, 1997. https://doi.org/10.3390/genes12121997
Berloco MF, Minervini CF, Moschetti R, Palazzo A, Viggiano L, Marsano RM. Evidence of the Physical Interaction between Rpl22 and the Transposable Element Doc5, a Heterochromatic Transposon of Drosophila melanogaster. Genes. 2021; 12(12):1997. https://doi.org/10.3390/genes12121997
Chicago/Turabian StyleBerloco, Maria Francesca, Crescenzio Francesco Minervini, Roberta Moschetti, Antonio Palazzo, Luigi Viggiano, and René Massimiliano Marsano. 2021. "Evidence of the Physical Interaction between Rpl22 and the Transposable Element Doc5, a Heterochromatic Transposon of Drosophila melanogaster" Genes 12, no. 12: 1997. https://doi.org/10.3390/genes12121997