KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
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| Mouse monoclonal anti-dimethyl-histone H3-K9 antibody | Abcam | ab1220; RRID:AB_449854 |
| Mouse monoclonal anti-c-Myc antibody | SIGMA | M4439; RRID:AB_439694 |
| Mouse monoclonal anti-FLAG M2 antibody | SIGMA | F1804; RRID:AB_262044 |
| Mouse anti-green fluorescent protein | Roche | 11814460001; RRID:AB_390913 |
| Mouse anti-HA monoclonal antibody | Sigma-Aldrich | 12CA5; RRID:AB_514505 |
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| Chemicals, Peptides, and Recombinant Proteins | ||
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| 5-fluoroorotic Acid (FOA) | USBiological | N/A |
| Thiabendazole | SIGMA | T8904 |
| EDTA-free Protease Inhibitor Cocktail | Roche | 11836170001 |
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| Critical Commercial Assays | ||
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| Ribo-Zero Gold rRNA Removal Kit | Illumina | MRZY1324 |
| TruSeq Stranded mRNA Library Prep Kit | Illumina | 20020594 |
| Agencourt AMPure XP beads | Beckman Coulter | A63880 |
| RNeasy MinElute Cleanup Kit | QIAGEN | 74204 |
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| Deposited Data | ||
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| RNaseq of iss1+ and isslΔC cells | SRA#PRJNA520927 | N/A |
| Hermes integration profiles | SRA#PRJNA517661 | N/A |
| ChIPseq of Iss1-3XFLAG | SRA#PRJNA517678 | N/A |
| ChIPseq of H3K9me2 | SRR10758214 SRR10758215 |
N/A |
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| Experimental Models: Organisms/Strains | ||
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| S. pombe YHL10155: mat1Msmt0 leu1-32 ade6-210 his2 ura4DS/E otr1R(Sph1)::ura4+ | (Chalamcharla et al., 2015) | N/A |
| S. pombe YHL12148: mat1Msmt0 leu1-32 ade6-216 his2 ura4 otr1R(Sph1)::ura4+ clr4Δ::KanMX | This paper | N/A |
| S. pombe: h+ leu1-32 ura4-D18 ade6-M216 clr4::KanMX | This paper | N/A |
| S. pombe YHL11414: h+ leu1-32 ura4-D18 ade6-M216 swi6::ura4 | This paper | N/A |
| S. pombe: h+ ade6-M216 ura4-D18 leu1-32 | Bioneer Corporation | ED668 |
| S. pombe YHL12159:h+ ade6-M216 ura4-D18 leu1-32 iss1ΔC-3xflag::KanMX | This study | N/A |
| S. pombe YHL12215:h+ ade6-M216 ura4-D18 leu1-32 iss1-3xflag::KanMX | This study | N/A |
| S. pombe YHL12160:mat1Msmt0 leu1-32 ade6-216 his2 ura4 otr1R(Sph1)::ura4+ iss1ΔC-3xflag::KanMX | This study | N/A |
| S. pombe YHL12266:h+ ade6 m210 his3-da leu1-32 ura4-d18 iss1-13xmyc::kanMX rrp6-3xflag::hyg | This study | N/A |
| S. pombe YHL12237:h+ leu1-32 ura4-D18 ade6-M216 rrp6-3xFlag::Hyg | This study | N/A |
| S. pombe YHL12783:YHL12237 transformed with pREP41-Iss1-HA | This study | N/A |
| S. pombe YHL12786:YHL12237 transformed with pREP41-Iss1ΔC HA | This study | N/A |
| S. pombe YHL12789:YHL12237 transformed with pREP41-GFP-HA | This study | N/A |
| S. pombe YHL12282:h+ ade6 m210 his3-da leu1-32 ura4-d18 mmi1-3xflag::hyg | This study | N/A |
| S. pombe YHL12792.YHL12282 transformed with pREP41-Iss1-HA | This study | N/A |
| S. pombe YHL12795.YHL12282 transformed with pREP41-Iss1ΔC HA | This study | N/A |
| S. pombe YHL12798.YHL12282 transformed with pREP41-GFP-HA | This study | N/A |
| S. pombe SR282 Paul Russell:h90 leu1-32 ade6-216 his2- ago1::KanMX6 | This study | N/A |
| S. pombe YHL12167:h90 leu1-32 ade6-216 his2- ago1::KanMX6 iss1ΔC-3xflag::KanMX | This study | N/A |
| S. pombe YHL12164:h+ leu1-32 ura4-D18 iss1 ΔC-3xflag::KanMX pRep41-N-GFP | This study | N/A |
| S. pombe YHL12162:h+ leu1-32 ura4-D18 iss1 ΔC-3xflag::KanMX pRep41-N-GFP-iss1 | This study | N/A |
| S. pombe YHL12179:h+ leu1-32 ura4-D18 pRep41-N-GFP | This study | N/A |
| S. pombe YHL12165:h+ leu1-32 ura4-D18 pRep41-N-GFP-iss1 | This study | N/A |
| S. pombe YHL12195:h+ ade6 m210 his3-da leu1-32 ura4-d18 iss1-GFP::KanMX | This study | N/A |
| S. pombe YHL12284 JT3 A. Yamashita:h90 pla1-GFP-kanR ade6-M216 leu1 | This study | N/A |
| S. pombe YHL12301:h90 pla1-GFP-kanR ade6-M216 leu1 pREP41-iss1 ΔC-3HA | This study | N/A |
| h90 pla1-GFP-kanR ade6-M216 leu1 pREP41-mCherry-iss1 | This study | N/A |
| S. pombe YHL12214:h+ ade6 m210 his3-da leu1-32 ura4-d18 iss1-13xmyc::kanMX | This study | N/A |
| S. pombe YHL12295:h+ ade6 m210 his3-da leu1-32 ura4-d18 iss1-13xmyc::kanMX mmi1-3xflag::hyg | This study | N/A |
| S. pombe YHL9451A:h- leu1-32, ura4-294 + pHL2577, pHL2578 | This study | N/A |
| S. pombe YHL9451C:h- leu1-32, ura4-294 + pHL2577, pHL2578 | This study | N/A |
| S. pombe YHL10161A:mat1Msmt0 leu1-32 ade6-210 his2 ura4DS/E otr1R(Sph1)::ura4+ pHL2577, pHL2578 | This study | N/A |
| S. pombe YHL10161B:mat1Msmt0 leu1-32 ade6-210 his2 ura4DS/E otr1R(Sph1)::ura4+ pHL2577, pHL2578 | This study | N/A |
| S. pombe YHL9451:YHL912 with pHL2577 | This study | N/A |
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| Oligonucleotides | ||
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| HL3835, forward primer for deletion of 38 aa from C-terminal of iss1, ATGCACAACCTACTCATAATCCTACATCGAGTTATGGGAATGGAGCCTCTACGAACTATAATGCCTCAAGACCTCCTAGTCGGATCCCCGGGTTAATTAA | This study | N/A |
| HL3836, reverse primer for deletion of 38 aa from C-terminal of iss1, TAGAAGTAGTCGACACCCCATAGCATACAATTAAAACGATTCTCCTAAGATTTGCATACACAAACGTTATTGTATTTATCGAATTCGAGCTCGTTTAAAC | This study | N/A |
| HL5086, forward primer for N-terminal tagging
of iss1, AAGGAATCCATAATTTCAGGTGCGAGACTCAGTCAATGA
ATGGACTTGCCCCATATACATTTTAGAAGTGG
GTTTTTACTGAATTCGAGCTCGTTTAAAC |
This study | N/A |
| HL5087, reverse primer for N-terminal tagging of iss1, ACTCCATCATTCCGTTCCTCTATGGTTTCTCCATATAAATATTCATCCTCATCAACATCCATGTGCTGATCAGCATTTGACATGATTTAACAAAGCGACTATA | This study | N/A |
| HL5041, forward primer for N-terminal tagging of mmi1, TGGAAATGATGTATGATGAAGGCAGCAGACTTTGTACGTTGATTAACTATGCTATAATGAAGCGAATTGGAAGAGACCGTCGGATCCCCGGGTTAATTAA | This study | N/A |
| HL5042, reverse primer for N-terminal tagging of mmi1, CCGCTCCAATAGTATATAGTTTCATGAATTTTGTTAAAGGAAGCATCCAACATTCTTGAAAATCCAGCAAAAGGGAGATAGAATTCGAGCTCGTTTAAAC | This study | N/A |
| HL4200, forward primer for N-terminal tagging of rrp6, TTAACAAGGTAAGTTTACCAAAGCGAAATACTAGGGAACTCAAAAAAAGGAAGGTTTCAGACGGGAAGAGTACATCTTATCGGATCCCCGGGTTAATTAA | This study | N/A |
| HL4197, reverse primer for N-terminal tagging of rrp6, TTTTCAATTTCTACAATGTATTTAAAAGTAAATTTTCATTTTGTCAAAATTGTTTCGTTTATTTAAAATTAAAACTGATAGAATTCGAGCTCGTTTAAAC | This study | N/A |
| HL1870, linker (adaptor) that ligates to MseI-digested end of genomic DNA fragment, GTAATACGACTCACTATAGGGCTCCGCTTAAGGGAC | (Guo et al., 2013) | N/A |
| HL1871, 5 Phos, 3 AmMO linker (adaptor) that ligates to MseI-digested end of genomic DNA fragment 5Phos/TAGTCCCTTAAGCGGAG/3AmMO/ | (Guo et al., 2013) | N/A |
| HL3504, The primer specific for the TIR end of Hermes with 5 bp barcode (ACGTC) at upstream, AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTACGTCCTATGTGGCTTACGTTTGCCTG | This study | N/A |
| HL3505, primer specific for the TIR end of Hermes with 5 bp barcode (TGCAG) at upstream, AATGATACGGCGACCACCGA GATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTTGCA GCTATGTGGCTTACGTTTGCCTG | This study | N/A |
| HL3506, primer specific for the TIR end of Hermes with 5 bp barcode (GATCA) at upstream, AATGATACGGCGACCACCGA GATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTGATC ACTATGTGGCTTACGTTTGCCTG | This study | N/A |
| HL3507, primer specific for the TIR end of Hermes with 5 bp barcode (CTAGT) at upstream, AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTCTAGTCTATGTGGCTTACGTTTGCCTG | This study | N/A |
| HL3508, primer specific for the TIR end of Hermes with 5 bp barcode (ACTGC) at upstream, AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTACTGCCTATGTGGCTTACGTTTGCCTG | This study | N/A |
| HL3509, primer specific to the adaptor used to amplify all Hermes libraries, CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCTGTAATACGACTCACTATAGG GC | This study’ | N/A |
| HL3643, primer specific for the TIR end of Hermes with 5 bp barcode (GTACG) at upstream, AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCGTACGTCTATGTGGCTTACGTTTGCCTG | This study | N/A |
| HL3644, primer specific for the TIR end of Hermes with | This study | N/A |
| HL3645, primer specific for the TIR end of Hermes with 5 bp barcode (CAGAT) at upstream, AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCCAGATTCTATGTGGCTTACGTTTGCCTG | This study | N/A |
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| Recombinant DNA | ||
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| pREP41-mCherry-iss1 | This study | N/A |
| pREP41-iss1ΔC-3HA, Akira Yamashita | This study | N/A |
| pREP41-iss1-3HA, Akira Yamashita | This study | N/A |
| pRep41-N-GFP-iss1 | This study | N/A |
| pRep41-N-GFP | This study | N/A |
| pHL2577: PSP2 containing kanMX6 flanked by Hermes left, right | (Guo et al., 2013) | N/A |
| pHL2578: Rep81×1 transposase (Hermes) express w/low nmt | (Guo et al., 2013) | N/A |
| pHL1768: Rep81X, Empty vector (serves as control, as there is no Hermes transposase) | (Guo et al., 2013) | N/A |
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| Software and Algorithms | ||
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| HTtools | (Esnault et al., 2019) | N/A |
| CLCbio Workbench v11 | QIAGEN | (https://digitalinsights.qiagen.com/) |
| Deeptools | (Ramirez et al., 2016) | https://deeptools.readthedocs.io/en/develop/content/list_of_tools.html |
| Lewis-Sigler Institute for Integrative Genomics web tool | (Boyle et al., 2004) | https://lsi.princeton.edu/research/facilities |