The Brm-HDAC3-Erm repressor complex suppresses dedifferentiation in Drosophila type II neuroblast lineages
- National University of Singapore, Singapore
- Duke-NUS Graduate Medical School Singapore, Singapore
- Institute for Research in Biomedicine, Spain
- Temasek Life Sciences Laboratory, Singapore
- Peter MacCallum Cancer Centre, Australia
- University of Melbourne, Australia
- University of Melbourne, Parkville, Australia
- Genome Institute of Singapore, Singapore
- Institució Catalana de Recerca i Estudis Avançats (ICREA), Spain
Figures
Figure 1 with 1 supplement
The Brm complex suppresses the formation of ectopic type II neuroblasts.
(A–C) Type II MARCM clones of control (the MARCM driver; D), brm2 (B, B′) and brmT362 (C, C′) were labeled with Dpn (blue), Ase (red) and CD8::GFP (green). (D) Quantification of neuroblast number …
Figure 1—figure supplement 1
Analysis of chromatin remodelers in larval brains.
(A and B) Larval brains of control (elav-Gal4 driver) and brm knockdown under the control of elav-Gal4 driver were labeled with Insc and Mira (A) and dMyc (B). Central brain is to the left of white …
Figure 2 with 1 supplement
The Brm complex suppresses INP dedifferentiation into type II neuroblasts.
(A–C′) Type II MARCM clones of control (the MARCM driver; A, A′), brm2 (B, B′) and brmT362 (C, C′) were labeled with Dpn (blue), Ase (red) and CD8::GFP (green). (D) Quantifications of INP number per …
Figure 2—figure supplement 1
Partial knock down of brm in INP clones.
Left panels, INP clones of a control (driver: erm-Gal4 [II]; erm-Gal4 [III]) and brm RNAi under erm-Gal4 (II); erm-Gal4 (III) with UAS-Dcr2 UAS-CD8-GFP were labeled with Brm (red) and CD8 (green). …
Figure 3 with 1 supplement
HDAC3 acts cooperatively with the Brm complex to suppress the formation of ectopic type II neuroblasts.
(A–D) The driver control (A), brm RNAi (B), hdac3 RNAi (C), brm hdac3 double knockdown (D) under the type II driver were labeled with Dpn, Ase, and CD8. (E) Quantification of neuroblast number per …
Figure 3—figure supplement 1
Brm is not important for the apical-basal polarity of neuroblasts.
(A–C) Neuroblast of control MARCM clones and brm2 MARCM clones were co-labeled with aPKC (white), CD8 (green) and Phospho-Histone H3 (PH3; green) (A) or Numb, GFP and DNA (B) or Brat, GFP and DNA (C)…
Figure 4 with 1 supplement
The Brm remodeling complex physically associates with Erm and HDAC3.
(A) Co-immunoprecipitation (Co-IP) between Flag-Brm and Myc-Erm. (B) An illustration of Erm domains and truncated constructs. (C) Co-IP between Myc-Brm and Flag-Erm-N or Flag-Erm-C. (D) Co-IP …
Figure 4—figure supplement 1
Brm and Erm may regulate gene expression of some common downstream targets.
(A) Co-IP was performed using S2 cells expressing Flag-PntP1 and Myc-Brm. IP was performed using anti-Flag or anti-Myc antibodies. Western blot was performed using anti-Flag and anti-Myc antibodies. …
Figure 5 with 1 supplement
Brm genetically interacts with Erm to prevent dedifferentiation of INPs to neuroblasts.
(A–D) Type II clones of control (the type II driver; A), brm knockdown (B), erm knockdown (C) and brm erm double knockdown (D) were labeled with Dpn (blue), Ase (red) and CD8 (green). (E) …
Figure 5—figure supplement 1
Knocking down of erm enhanced the neuroblast overgrowth observed in brm mutants.
(A) Type II MARCM clones of brmT362 and UAS-Dcr2; erm RNAi, brmT362 were labeled with Dpn, Ase and CD8. Arrows, neuroblasts. Clone outline is indicated by white dotted line. Scale bar, 10 µm. (B) …
Tables
Table 1
Histone modifiers and their RNAi lines
| S/No. | Gene name | Full name | CG # | Main function | VDRC RNAi lines |
|---|---|---|---|---|---|
| 1 | enok | Enoki mmushroom | CG11290 | HAT | KK108400, GD37527 |
| 2 | nej | Nejire/CBP | CG15319 | HAT | KK105115 |
| 3 | CG1894 | CG1894 | HAT | GD41575, GD41574 | |
| 4 | CG2051 | CG2051 | HAT | GD33458 | |
| 5 | Mof | Males absent on the first | CG3025 | HAT | KK105370 |
| 6 | Rpb4 | Rpb4 | CG33520 | HAT | GD21985, GD23308 |
| 7 | Pcaf | Gcn | CG4107 | HAT | KK108943, GD21786 |
| 8 | YL-1 | YL-1 | CG4621 | HAT | GD21903 |
| 9 | Chm | Chameau | CG5229 | HAT | KK105542 |
| 10 | Dik | Diskette | CG7098 | HAT | GD46320 |
| 11 | lid | Little imaginal discs | CG9088 | HAT | GD42203, KK103830 |
| 12 | Ada2b | CG9638 | HAT | GD24076 | |
| 13 | Sirt7 | CG11305 | HDAC | GD18043, GD18045 | |
| 14 | HDAC4 | CG1770 | HDAC | GD20522 | |
| 15 | HDAC3 | CG2128 | HDAC | KK107073 | |
| 16 | HDACX | CG31119 | HDAC | KK108098 | |
| 17 | Sirt4 | CG3187 | HDAC | GD40295, KK110639 | |
| 18 | Sirt2 | CG5085 | HDAC | KK103790 | |
| 19 | Sir2 | CG5216 | HDAC | GD23199, KK108241, KK105502 | |
| 20 | Bin1 | Bicoid interacting protein | CG6046 | HDAC | KK105352, GD15710 |
| 21 | Sirt6 | CG6284 | HDAC | GD22483 | |
| 22 | Gug | Grunge | CG6964 | HDAC | GD13687 |
| 23 | Rpd3 | HDAC1 | CG7471 | HDAC | GD46929, GD30600, GD46929 |
| 24 | Sin3a | CG8815 | HDAC | KK105852 | |
| 25 | Rtf1 | CG10955 | Methyl transferase | KK110392 | |
| 26 | Vig2 | CG11844 | Methyl transferase | KK107081, GD17245 | |
| 27 | egg | eggless | CG12196 | Methyl transferase | KK101677, GD33730 |
| 28 | esc | extra sexcombs | CG14941 | Methyl transferase | GD5690, GD5692 |
| 29 | set2 | CG1716 | Methyl transferase | GD30707 | |
| 30 | g9a | CG2995 | Methyl transferase | GD25474 | |
| 31 | pr-set7 | CG3307 | Methyl transferase | KK105422 | |
| 32 | trr | trithorax-related | CG3848 | Methyl transferase | GD10749, KK110276 |
| 33 | CG40351 | CG40351 | Methyl transferase | GD40683, GD10833, GD45267 | |
| 34 | CG4565 | CG4565 | Methyl transferase | GD5665 | |
| 35 | mes-4 | CG4976 | Methyl transferase | GD10836 | |
| 36 | Art4 | Arginine methyl transferase 4 | CG5358 | Methyl transferase | KK107009 |
| 37 | Su(var)3–9 | auppressor of variegation 3–9 | CG6476 | Methyl transferase | GD39377 |
| 38 | Art1 | Arginine methyl transferase 11 | CG6554 | Methyl transferase | GD40388, KK110391 |
| 39 | ash2 | absent, small or homeotic discs 2 | CG6677 | Methyl transferase | KK100718 |
| 40 | LKR | Lysine ketoglutarate reductase | CG7144 | Methyl transferase | GD51346 |
| 41 | Su(z)12 | Suppressor of Zeste 205 | CG8013 | Methyl transferase | GD42422, GD42423 |
| 42 | Su(var)205 | Suppressor of variegation 205 | CG8409 | Methyl transferase | KK107477 |
| 43 | Ash1 | Absent, small or homeotic discs 1 | CG8887 | Methyl transferase | GD28928 |
Table 2
Predicted common target genes of Brm and Erm
| S/No. | CG name | Gene name |
|---|---|---|
| 1 | CG10033 | for |
| 2 | CG10117 | ttv |
| 3 | CG10137 | CG10137 |
| 4 | CG10159 | BEAF-32 |
| 5 | CG10388 | Ubx |
| 6 | CG10610 | ECSIT |
| 7 | CG1071 | E2f2 |
| 8 | CG10844 | RyR |
| 9 | CG1100 | Rpn5 |
| 10 | CG11228 | hpo |
| 11 | CG11309 | CG11309 |
| 12 | CG11589 | VhaM9.7-c |
| 13 | CG12165 | Incenp |
| 14 | CG12321 | CG12321 |
| 15 | CG12333 | CG12333 |
| 16 | CG12387 | zetaTry |
| 17 | CG12797 | Ciao1 |
| 18 | CG12818 | CG12818 |
| 19 | CG12819 | sle |
| 20 | CG12855 | HPS1 |
| 21 | CG12994 | CG12994 |
| 22 | CG13004 | CG13004 |
| 23 | CG13016 | CG13016 |
| 24 | CG13018 | CG13018 |
| 25 | CG13117 | CG13117 |
| 26 | CG1322 | zfh1 |
| 27 | CG13316 | Mnt |
| 28 | CG13350 | Ctf4 |
| 29 | CG13366 | CG13366 |
| 30 | CG13432 | qsm |
| 31 | CG13472 | CG13472 |
| 32 | CG13688 | Ipk2 |
| 33 | CG13900 | CG13900 |
| 34 | CG13919 | CG13919 |
| 35 | CG14291 | CG14291 |
| 36 | CG14463 | CG14463 |
| 37 | CG1453 | Klp10A |
| 38 | CG14813 | deltaCOP |
| 39 | CG14814 | CG14814 |
| 40 | CG14938 | crol |
| 41 | CG14939 | CycY |
| 42 | CG15010 | ago |
| 43 | CG15027 | CG15027 |
| 44 | CG15120 | CG15120 |
| 45 | CG15387 | CG15387 |
| 46 | CG15701 | CG15701 |
| 47 | CG15706 | CG15706 |
| 48 | CG15845 | Adf1 |
| 49 | CG1600 | Drat |
| 50 | CG1616 | dpa |
| 51 | CG17033 | elgi |
| 52 | CG17035 | GXIVsPLA2 |
| 53 | CG17052 | obst-A |
| 54 | CG17233 | CG17233 |
| 55 | CG17249 | CG17249 |
| 56 | CG17259 | CG17259 |
| 57 | CG17260 | CG17260 |
| 58 | CG1765 | EcR |
| 59 | CG17803 | CG17803 |
| 60 | CG1785 | CG1785 |
| 61 | CG1817 | Ptp10D |
| 62 | CG18292 | CG18292 |
| 63 | CG1845 | Br140 |
| 64 | CG18660 | Nckx30C |
| 65 | CG18675 | CG18675 |
| 66 | CG2004 | CG2004 |
| 67 | CG2019 | disp |
| 68 | CG2051 | CG2051 |
| 69 | CG2146 | didum |
| 70 | CG2189 | Dfd |
| 71 | CG2446 | Amun |
| 72 | CG2698 | CG2698 |
| 73 | CG2720 | Hop |
| 74 | CG2813 | cold |
| 75 | CG2977 | Inx7 |
| 76 | CG3059 | NTPase |
| 77 | CG3127 | Pgk |
| 78 | CG31481 | pb |
| 79 | CG3157 | gammaTub23C |
| 80 | CG3165 | CG3165 |
| 81 | CG3166 | aop |
| 82 | CG31712 | CG31712 |
| 83 | CG31713 | Apf |
| 84 | CG3178 | Rrp1 |
| 85 | CG31794 | Pax |
| 86 | CG31852 | Tap42 |
| 87 | CG31855 | CG31855 |
| 88 | CG31911 | Ent2 |
| 89 | CG32022 | CG32022 |
| 90 | CG32556 | chas |
| 91 | CG32592 | hiw |
| 92 | CG33116 | CG33116 |
| 93 | CG33162 | SrpRbeta |
| 94 | CG3587 | CG3587 |
| 95 | CG3666 | Tsf3 |
| 96 | CG3842 | CG3842 |
| 97 | CG3857 | CG3857 |
| 98 | CG3920 | Reph |
| 99 | CG42254 | CG42254 |
| 100 | CG42311 | grh |
| 101 | CG42334 | comm3 |
| 102 | CG42362 | CG42362 |
| 103 | CG42363 | CG42363 |
| 104 | CG42365 | CG42365 |
| 105 | CG42379 | CG42379 |
| 106 | CG42380 | CG42380 |
| 107 | CG42381 | CG42381 |
| 108 | CG4400 | CG4400 |
| 109 | CG4590 | Inx2 |
| 110 | CG4619 | CG4619 |
| 111 | CG4645 | CG4645 |
| 112 | CG4798 | l(2)k01209 |
| 113 | CG4996 | CG4996 |
| 114 | CG5229 | chm |
| 115 | CG5393 | apt |
| 116 | CG5505 | scny |
| 117 | CG5548 | CG5548 |
| 118 | CG5588 | Mtl |
| 119 | CG5599 | CG5599 |
| 120 | CG5611 | CG5611 |
| 121 | CG5613 | CG5613 |
| 122 | CG5824 | l(3)07882 |
| 123 | CG5836 | SF1 |
| 124 | CG6022 | Cchl |
| 125 | CG6202 | Surf4 |
| 126 | CG6218 | CG6218 |
| 127 | CG6235 | tws |
| 128 | CG6241 | CG6241 |
| 129 | CG6272 | CG6272 |
| 130 | CG6322 | U4-U6-60K |
| 131 | CG6343 | ND42 |
| 132 | CG6401 | CG6401 |
| 133 | CG6511 | CG6511 |
| 134 | CG6556 | cnk |
| 135 | CG6565 | CG6565 |
| 136 | CG6604 | H15 |
| 137 | CG6634 | mid |
| 138 | CG6829 | Ark |
| 139 | CG6948 | Clc |
| 140 | CG6951 | CG6951 |
| 141 | CG6983 | CG6983 |
| 142 | CG7082 | papi |
| 143 | CG7085 | l(2)s5379 |
| 144 | CG7186 | SAK |
| 145 | CG7191 | CG7191 |
| 146 | CG7372 | CG7372 |
| 147 | CG7379 | CG7379 |
| 148 | CG7564 | CG7564 |
| 149 | CG7597 | Cdk12 |
| 150 | CG7632 | CG7632 |
| 151 | CG7685 | CG7685 |
| 152 | CG7734 | shn |
| 153 | CG7771 | sim |
| 154 | CG7828 | APP-BP1 |
| 155 | CG7845 | CG7845 |
| 156 | CG7849 | CG7849 |
| 157 | CG7957 | MED17 |
| 158 | CG7961 | alphaCop |
| 159 | CG8067 | CG8067 |
| 160 | CG8241 | pea |
| 161 | CG8287 | Rab8 |
| 162 | CG8360 | CG8360 |
| 163 | CG8372 | CG8372 |
| 164 | CG8396 | Ssb-c31a |
| 165 | CG8409 | Su(var)205 |
| 166 | CG8481 | CG8481 |
| 167 | CG8790 | Dic1 |
| 168 | CG8798 | Lon |
| 169 | CG8817 | lilli |
| 170 | CG9042 | Gpdh |
| 171 | CG9054 | Ddx1 |
| 172 | CG9063 | Rich |
| 173 | CG9065 | CG9065 |
| 174 | CG9243 | CG43345 |
| 175 | CG9243 | CG43346 |
| 176 | CG9244 | Acon |
| 177 | CG9249 | CG9249 |
| 178 | CG9250 | Mpp6 |
| 179 | CG9305 | CG9305 |
| 180 | CG9376 | CG9376 |
| 181 | CG9473 | MED6 |
| 182 | CG9596 | CG9596 |
| 183 | CG9635 | RhoGEF2 |
| 184 | CG9641 | CG9641 |
| 185 | CG9730 | mRpL21 |
| 186 | CG9750 | rept |
| 187 | CG9829 | poly |
| 188 | CG9865 | CG9865 |
