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Laura V. Glaser, Simone Rieger, Sybille Thumann, Sophie Beer, Cornelia Kuklik-Roos, Dietmar E. Martin, Kerstin C. Maier, Marie L. Harth-Hertle, Björn Grüning, Rolf Backofen, Stefan Krebs, Helmut Blum, Ralf Zimmer, Florian Erhard, Bettina Kempkes
EBF1 binds to EBNA2 and promotes the assembly of EBNA2 chromatin complexes in B cells
www.plospathogens.org
www.plospathogens.org
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(a)Tj
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3.2769 0 Td
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(18)Tj
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1.7404 0 Td
(19)Tj
0 g
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(EBNA2)Tj
3.2598 0 Td
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(histone)Tj
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(3)Tj
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(lysine)Tj
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(27)Tj
1.1735 0 Td
(acetylation)Tj
4.5581 0 Td
(\(H3K27ac\))Tj
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(and)Tj
1.7348 0 Td
(H3K4)Tj
2.6135 0 Td
(mono-methylation)Tj
7.7725 0 Td
(\(H3K4me1\))Tj
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(11)Tj
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(20)Tj
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(EBNA2)Tj
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(PU.1/SPI1)Tj
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(EBF1)Tj
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1.1849 0 Td
(modulating)Tj
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(the)Tj
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(chromatin)Tj
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(state)Tj
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(and)Tj
1.7291 0 Td
(thereby)Tj
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(promoting)Tj
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(access)Tj
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(of)Tj
1.0148 0 Td
(EBNA2)Tj
3.2655 0 Td
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(chromatin,)Tj
4.6431 0 Td
(indirectly.)Tj
4.2746 0 Td
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5.5955 0 Td
(EBNA2)Tj
3.2655 0 Td
(co-occurring)Tj
5.4028 0 Td
(factors)Tj
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(might)Tj
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(serve)Tj
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(as)Tj
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(alternate)Tj
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(adaptors)Tj
3.668 0 Td
(that)Tj
1.7801 0 Td
(promote)Tj
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(DNA)Tj
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(binding)Tj
3.3562 0 Td
(of)Tj
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(EBNA2.)Tj
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(CBF1)Tj
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(is)Tj
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(ubiquitously)Tj
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4.0932 0 Td
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(cells)Tj
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(including)Tj
4.0422 0 Td
(primary)Tj
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(human)Tj
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(B)Tj
0.7994 0 Td
(cells)Tj
-34.3443 -1.3039 Td
(and)Tj
1.7291 0 Td
(EBV)Tj
2.0636 0 Td
(infected)Tj
3.4185 0 Td
(and)Tj
1.7348 0 Td
(non-infected)Tj
5.3745 0 Td
(human)Tj
3.0727 0 Td
(B)Tj
0.8051 0 Td
(cell)Tj
1.559 0 Td
(lines.)Tj
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1.6271 0 Td
(this)Tj
1.6724 0 Td
(study,)Tj
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(we)Tj
1.3266 0 Td
(used)Tj
2.0636 0 Td
(a)Tj
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(CBF1)Tj
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(defi-)Tj
-34.469 -1.2983 Td
(cient)Tj
2.1769 0 Td
(human)Tj
3.0784 0 Td
(B)Tj
0.7994 0 Td
(cell)Tj
1.5647 0 Td
(line,)Tj
1.9276 0 Td
(which)Tj
2.6532 0 Td
(had)Tj
1.7064 0 Td
(been)Tj
2.1203 0 Td
(generated)Tj
4.1386 0 Td
(by)Tj
1.1792 0 Td
(homologous)Tj
5.1987 0 Td
(recombination)Tj
6.1398 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
-33.7207 -1.2982 Td
(somatic)Tj
3.3278 0 Td
(B)Tj
0.7994 0 Td
(cell)Tj
1.559 0 Td
(line)Tj
1.7065 0 Td
(DG75,)Tj
2.8516 0 Td
(to)Tj
1.0205 0 Td
(screen)Tj
2.7722 0 Td
(for)Tj
1.389 0 Td
(CBF1)Tj
2.4661 0 Td
(independent)Tj
5.2724 0 Td
(functions)Tj
3.9912 0 Td
(of)Tj
1.0148 0 Td
(EBNA2.)Tj
3.4922 0 Td
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1.7802 0 Td
(parental)Tj
-33.4429 -1.2983 Td
(DG75)Tj
2.6191 0 Td
(B)Tj
0.8051 0 Td
(cell)Tj
1.559 0 Td
(line)Tj
1.7008 0 Td
(is)Tj
0.8504 0 Td
(an)Tj
1.2019 0 Td
(EBV)Tj
2.0636 0 Td
(negative)Tj
3.5376 0 Td
(Burkitt's)Tj
3.634 0 Td
(lymphoma)Tj
4.5467 0 Td
(cell)Tj
1.5591 0 Td
(line.)Tj
1.9275 0 Td
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2.6532 0 Td
(the)Tj
1.4684 0 Td
(knock-down)Tj
5.3121 0 Td
(of)Tj
-35.4385 -1.3039 Td
(EBF1)Tj
2.3697 0 Td
(and)Tj
1.7348 0 Td
(CBF1)Tj
2.4661 0 Td
(in)Tj
1.0318 0 Td
(EBV)Tj
2.0636 0 Td
(immortalized)Tj
5.6183 0 Td
(B)Tj
0.7993 0 Td
(cells)Tj
1.9219 0 Td
(severely)Tj
3.3732 0 Td
(impairs)Tj
3.2598 0 Td
(cellular)Tj
3.1408 0 Td
(viability)Tj
3.4129 0 Td
([)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(],)Tj
2.0806 0 Td
(DG75)Tj
2.6192 0 Td
(cells)Tj
-35.892 -1.2983 Td
(tolerate)Tj
3.2201 0 Td
(inactivation)Tj
4.9663 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(CBF1)Tj
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(gene)Tj
2.0806 0 Td
(without)Tj
3.3279 0 Td
(loss)Tj
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(of)Tj
1.0148 0 Td
(viability)Tj
3.4185 0 Td
([)Tj
0.83 0.64 0.02 0 k
(21)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.7348 0 Td
(22)Tj
0 g
(].)Tj
1.7348 0 Td
(We)Tj
1.6101 0 Td
(compared)Tj
4.2462 0 Td
(EBNA2)Tj
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(induced)Tj
3.4639 0 Td
(cellular)Tj
3.1407 0 Td
(genes)Tj
2.4378 0 Td
(in)Tj
1.0375 0 Td
(CBF1)Tj
2.4661 0 Td
(proficient)Tj
4.1329 0 Td
(and)Tj
1.7291 0 Td
(deficient)Tj
3.6907 0 Td
(DG75)Tj
2.6192 0 Td
(cells)Tj
1.9219 0 Td
(and)Tj
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(found)Tj
2.6192 0 Td
(the)Tj
1.4683 0 Td
(majority)Tj
3.6284 0 Td
(of)Tj
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(EBNA2)Tj
3.2654 0 Td
(target)Tj
2.5115 0 Td
(genes)Tj
2.4378 0 Td
(to)Tj
1.0261 0 Td
(be)Tj
1.1509 0 Td
(CBF1)Tj
2.4661 0 Td
(dependent.)Tj
4.6828 0 Td
(A)Tj
0.9071 0 Td
(minor)Tj
2.7326 0 Td
(fraction)Tj
3.3562 0 Td
(of)Tj
1.0148 0 Td
(EBNA2)Tj
3.2655 0 Td
(target)Tj
2.5115 0 Td
(genes)Tj
2.4434 0 Td
(is)Tj
0.8447 0 Td
(regu-)Tj
-34.6164 -1.2982 Td
(lated)Tj
2.1486 0 Td
(CBF1)Tj
2.4661 0 Td
(independently.)Tj
6.2022 0 Td
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1.2529 0 Td
(chromatin)Tj
4.4163 0 Td
(immunoprecipitation)Tj
8.8498 0 Td
(and)Tj
1.7347 0 Td
(genome)Tj
3.4073 0 Td
(wide)Tj
2.1203 0 Td
(sequencing)Tj
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(of)Tj
1.0147 0 Td
(EBNA2)Tj
3.2655 0 Td
(bound)Tj
2.8347 0 Td
(DNA)Tj
2.3811 0 Td
(fragments)Tj
4.2292 0 Td
(\(ChIP-Seq\),)Tj
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(we)Tj
1.3266 0 Td
(identified)Tj
4.0591 0 Td
(a)Tj
0.652 0 Td
(subpopulation)Tj
5.9867 0 Td
(of)Tj
1.0148 0 Td
(CBF1)Tj
2.4662 0 Td
(inde-)Tj
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(pendent)Tj
3.5035 0 Td
(EBNA2)Tj
3.2655 0 Td
(binding)Tj
3.3562 0 Td
(sites)Tj
1.9276 0 Td
(that)Tj
1.7801 0 Td
(was)Tj
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(significantly)Tj
5.0853 0 Td
(enriched)Tj
3.7361 0 Td
(for)Tj
1.3833 0 Td
(EBF1)Tj
2.3754 0 Td
(binding)Tj
3.3505 0 Td
(motifs.)Tj
2.9877 0 Td
(We)Tj
-34.452 -1.2983 Td
(show)Tj
2.296 0 Td
(that)Tj
1.7801 0 Td
(CBF1)Tj
2.4662 0 Td
(independent)Tj
5.278 0 Td
(EBNA2)Tj
3.2655 0 Td
(binding)Tj
3.3506 0 Td
(to)Tj
1.0261 0 Td
(chromatin)Tj
4.4164 0 Td
(is)Tj
0.8447 0 Td
(dependent)Tj
4.4617 0 Td
(on)Tj
1.2756 0 Td
(EBF1)Tj
2.3697 0 Td
(protein)Tj
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(expression.)Tj
4.6884 0 Td
(Importantly,)Tj
5.2838 0 Td
(we)Tj
1.3266 0 Td
(demonstrate)Tj
5.2327 0 Td
(that)Tj
1.7801 0 Td
(EBNA2)Tj
3.2655 0 Td
(and)Tj
1.7292 0 Td
(EBF1)Tj
2.3697 0 Td
(can)Tj
1.6271 0 Td
(form)Tj
2.2053 0 Td
(protein)Tj
3.1521 0 Td
(complexes)Tj
-32.6605 -1.2982 Td
(in)Tj
1.0318 0 Td
(CBF1)Tj
2.4717 0 Td
(positive)Tj
3.3109 0 Td
(and)Tj
1.7348 0 Td
(negative)Tj
3.5319 0 Td
(cells,)Tj
2.1487 0 Td
(indicating)Tj
4.2633 0 Td
(that)Tj
1.7801 0 Td
(EBF1)Tj
2.3754 0 Td
(serves)Tj
2.6136 0 Td
(as)Tj
1.0147 0 Td
(B)Tj
0.7994 0 Td
(cell)Tj
1.5591 0 Td
(specific)Tj
3.1918 0 Td
(DNA)Tj
2.3754 0 Td
(anchor)Tj
-34.2026 -1.3039 Td
(for)Tj
1.3832 0 Td
(EBNA2.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 314.022 cm
BT
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1 TL
-0.0033 Tc
0 0 Td
(Results)Tj
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0 -1.5023 Td
(Genome)Tj
4.0205 0 Td
(wide)Tj
2.2441 0 Td
(expression)Tj
4.9937 0 Td
(profiling)Tj
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(identifies)Tj
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(transcripts)Tj
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(regulated)Tj
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1.2614 0 Td
(EBNA2)Tj
3.4724 0 Td
(in)Tj
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(CBF1)Tj
2.7591 0 Td
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3.893 0 Td
(B)Tj
0.8834 0 Td
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Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 264.9826 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
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1.1055 0 Td
(order)Tj
2.4207 0 Td
(to)Tj
1.0262 0 Td
(rigorously)Tj
4.2973 0 Td
(test)Tj
1.6044 0 Td
(if)Tj
0.7766 0 Td
(EBNA2)Tj
3.2655 0 Td
(can)Tj
1.6214 0 Td
(exert)Tj
2.2054 0 Td
(any)Tj
1.6554 0 Td
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(the)Tj
1.474 0 Td
(absence)Tj
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1.1452 0 Td
(DNA)Tj
2.3754 0 Td
(adaptor)Tj
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(CBF1,)Tj
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(a)Tj
0.652 0 Td
(microarray)Tj
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(based)Tj
2.4774 0 Td
(genome)Tj
3.4073 0 Td
(wide)Tj
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(screen)Tj
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(EBNA2)Tj
3.2655 0 Td
(target)Tj
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(genes)Tj
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(in)Tj
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(DG75)Tj
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(B)Tj
0.7994 0 Td
(cells)Tj
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(that)Tj
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(either)Tj
2.5342 0 Td
(proficient)Tj
4.1329 0 Td
(\(wt\))Tj
1.8821 0 Td
(or)Tj
1.0942 0 Td
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3.6907 0 Td
(\(ko\))Tj
1.9049 0 Td
(for)Tj
1.3833 0 Td
(CBF1)Tj
2.4661 0 Td
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1.7008 0 Td
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4.4673 0 Td
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(Fig)Tj
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(1A,)Tj
1.6044 0 Td
(1B)Tj
1.2756 0 Td
(and)Tj
1.7347 0 Td
(1F)Tj
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1.2189 0 Td
(and)Tj
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1.7348 0 Td
(S1)Tj
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(\).)Tj
3.0047 0 Td
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2.1316 0 Td
(cell)Tj
1.5647 0 Td
(lines)Tj
2.0636 0 Td
(constitutively)Tj
5.5956 0 Td
(express)Tj
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(an)Tj
1.2076 0 Td
(estrogen)Tj
3.617 0 Td
(receptor)Tj
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(\(ER\))Tj
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(hormone)Tj
3.9231 0 Td
(binding)Tj
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(domain)Tj
3.3221 0 Td
(EBNA2)Tj
3.2655 0 Td
(fusion)Tj
2.7213 0 Td
(protein)Tj
3.1521 0 Td
(\(ER/EBNA2\).)Tj
5.6919 0 Td
(ER/EBNA2)Tj
4.7849 0 Td
(is)Tj
0.8503 0 Td
(retained)Tj
3.5093 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(cytoplasm)Tj
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(of)Tj
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1.559 0 Td
(but)Tj
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(rapidly)Tj
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(activated)Tj
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(and)Tj
1.7291 0 Td
(translocated)Tj
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(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(nucleus)Tj
3.2768 0 Td
(in)Tj
1.0375 0 Td
(response)Tj
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(to)Tj
1.0262 0 Td
(estrogen)Tj
3.617 0 Td
([)Tj
0.83 0.64 0.02 0 k
(21)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.7404 0 Td
(22)Tj
0 g
(].)Tj
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1.627 0 Td
(expression)Tj
4.4617 0 Td
(profiling,)Tj
3.9232 0 Td
(DG75)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 324.17 191.4519 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(ER/EBNA2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 360.5669 186.9732 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CBF1)Tj
2.4661 0 Td
(wt)Tj
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(and)Tj
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(CBF1)Tj
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(ko)Tj
1.2246 0 Td
(cells)Tj
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(were)Tj
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(cultured)Tj
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(in)Tj
1.0375 0 Td
(estrogen)Tj
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(supplemented)Tj
5.845 0 Td
(media)Tj
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(for)Tj
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