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Raphael Souza Pavani, Marcelo Santos da Silva, Carlos Alexandre Henrique Fernandes, Flavia Souza Morini, Christiane Bezerra Araujo, Marcos Roberto de Mattos Fontes, Osvaldo Augusto Sant’Anna, Carlos Renato Machado, Maria Isabel Cano, Stenio Perdigão Fragoso, Maria Carolina Elias
Replication Protein A Presents Canonical Functions and Is Also Involved in the Differentiation Capacity of Trypanosoma cruzi
www.plosntds.org
www.plosntds.org
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(\(CDK1\)/)Tj
3.7757 0 Td
(cyclin)Tj
2.5852 0 Td
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([)Tj
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(13)Tj
0 g
(])Tj
1.8539 0 Td
([)Tj
0.83 0.64 0.02 0 k
(14)Tj
0 g
(])Tj
1.8482 0 Td
(and)Tj
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(in)Tj
1.0374 0 Td
(response)Tj
3.7304 0 Td
(to)Tj
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(DNA)Tj
2.381 0 Td
(damage,)Tj
3.549 0 Td
(when)Tj
2.4038 0 Td
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(ATM)Tj
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(\(ataxia)Tj
2.8913 0 Td
(telangiectasia)Tj
5.5446 0 Td
(mutated\),)Tj
4.1102 0 Td
(ATR)Tj
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(\(ATM)Tj
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7.3077 0 Td
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3.1521 0 Td
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3.0897 0 Td
([)Tj
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(15)Tj
0 g
(].)Tj
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(RPA-3)Tj
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-34.401 -1.2983 Td
(thought)Tj
3.3732 0 Td
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1.0204 0 Td
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3.4413 0 Td
(the)Tj
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(RPA)Tj
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(16)Tj
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(trypanosomatids,)Tj
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(protozoa)Tj
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(that)Tj
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(appear)Tj
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(early)Tj
2.1544 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.4683 0 Td
(evolution)Tj
4.0025 0 Td
(of)Tj
1.0148 0 Td
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1.1055 0 Td
(1992,)Tj
2.347 0 Td
(Brown)Tj
2.914 0 Td
(and)Tj
1.7292 0 Td
(collaborators)Tj
5.4311 0 Td
(purified)Tj
3.4129 0 Td
(the)Tj
-34.2706 -1.3039 Td
(RPA)Tj
2.0862 0 Td
(complex)Tj
3.617 0 Td
(from)Tj
/F14 1 Tf
2.211 0 Td
[()13()-9(\r)14()10()18(\r)4()12(\r)4()]TJ
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[()9()-3(\n)1(\r)4(\n)1()4()4()22()11()]TJ
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4.3483 0 Td
(and)Tj
1.7291 0 Td
(demonstrated)Tj
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(that)Tj
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(a)Tj
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(nuclear)Tj
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(protein)Tj
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(can)Tj
1.6157 0 Td
(stimulate)Tj
3.8834 0 Td
(DNA)Tj
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(pol)Tj
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()Tj
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2.0466 0 Td
(but)Tj
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(cannot)Tj
2.965 0 Td
(replace)Tj
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(human)Tj
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(RPA)Tj
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(in)Tj
1.0205 0 Td
(the)Tj
1.4627 0 Td
(SV40)Tj
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(DNA)Tj
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(replication)Tj
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(reaction)Tj
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([)Tj
0.83 0.64 0.02 0 k
(17)Tj
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(].)Tj
2.0749 0 Td
(In)Tj
/F14 1 Tf
1.1112 0 Td
[()-3()2(\r)-4()12()18( )6()16()18(\r)4()]TJ
4.7848 0 Td
[()16( )6()-3()11()6()2()10(\r)-4()]TJ
/F9 1 Tf
(,)Tj
5.4482 0 Td
(LaRPA-1)Tj
3.8834 0 Td
(has)Tj
1.5421 0 Td
(relevant)Tj
3.4242 0 Td
(structural)Tj
4.0762 0 Td
(differences)Tj
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(compared)Tj
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(with)Tj
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(RPA-1)Tj
2.9083 0 Td
(from)Tj
2.2053 0 Td
(humans)Tj
3.4413 0 Td
(and)Tj
1.7291 0 Td
(yeast)Tj
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([)Tj
0.83 0.64 0.02 0 k
(18)Tj
0 g
(].)Tj
2.0749 0 Td
(In)Tj
/F14 1 Tf
1.1112 0 Td
[()-3()2(\r)-4()12()18( )6()16()18(\r)4()]TJ
4.7849 0 Td
[( )6()16()3()11()]TJ
/F9 1 Tf
(,)Tj
2.7949 0 Td
(LmRPA-1)Tj
4.2689 0 Td
(accumulates)Tj
5.1364 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(DNA)Tj
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(and)Tj
1.7291 0 Td
(co-localizes)Tj
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(with)Tj
1.9956 0 Td
(an)Tj
1.2019 0 Td
(LmHus1)Tj
3.7077 0 Td
(homologue)Tj
4.7508 0 Td
(after)Tj
2.0353 0 Td
(treatment)Tj
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(with)Tj
1.9956 0 Td
(hydroxyurea)Tj
5.2837 0 Td
(\(HU\))Tj
2.4038 0 Td
(and)Tj
-34.0665 -1.2983 Td
(camptothecin,)Tj
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(but)Tj
1.5534 0 Td
(the)Tj
1.474 0 Td
(interaction)Tj
4.6034 0 Td
(of)Tj
1.0148 0 Td
(these)Tj
2.2564 0 Td
(two)Tj
1.7064 0 Td
(proteins)Tj
3.515 0 Td
(has)Tj
1.542 0 Td
(not)Tj
1.576 0 Td
(been)Tj
2.1203 0 Td
(confirmed,)Tj
4.6262 0 Td
(so)Tj
1.0828 0 Td
(trypanoso-)Tj
-33.0007 -1.3039 Td
(matids)Tj
2.9253 0 Td
(RPA)Tj
2.0863 0 Td
(involvement)Tj
5.21 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(repair/damage)Tj
6.0378 0 Td
(response)Tj
3.736 0 Td
(remains)Tj
3.4413 0 Td
(obscure)Tj
3.3448 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
2.075 0 Td
(Despite)Tj
3.2485 0 Td
(this)Tj
-34.6108 -1.2983 Td
(evidence,)Tj
3.9344 0 Td
(in)Tj
/F14 1 Tf
1.0375 0 Td
[()-3()2(\r)-4()12()18( )6()16()18(\r)4()]TJ
/F9 1 Tf
(,)Tj
5.0116 0 Td
(only)Tj
1.9786 0 Td
(RPA-1)Tj
2.9083 0 Td
(has)Tj
1.5477 0 Td
(been)Tj
2.1203 0 Td
(studied)Tj
3.1521 0 Td
(while)Tj
2.3698 0 Td
(RPA)Tj
2.0863 0 Td
(from)Tj
/F14 1 Tf
2.2053 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
/F14 1 Tf
1.0318 0 Td
[(\n)1()-9(\r)]TJ
/F9 1 Tf
2.1657 0 Td
(and)Tj
/F14 1 Tf
1.7291 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
/F14 1 Tf
1.0318 0 Td
[()11()-9()4(\n)7()2(\r)]TJ
/F9 1 Tf
-34.3103 -1.2983 Td
(have)Tj
2.0579 0 Td
(never)Tj
2.4434 0 Td
(been)Tj
2.1203 0 Td
(described.)Tj
4.2803 0 Td
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2.4321 0 Td
(we)Tj
1.3266 0 Td
(first)Tj
1.8029 0 Td
(cloned,)Tj
3.1237 0 Td
(expressed)Tj
4.0932 0 Td
(and)Tj
1.7292 0 Td
(purified)Tj
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(RPA-1)Tj
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(and)Tj
1.7291 0 Td
(RPA-2)Tj
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(from)Tj
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2.2053 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
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1.0375 0 Td
[(\n)1()-9(\r)]TJ
/F9 1 Tf
2.16 0 Td
(\(respectively)Tj
5.2213 0 Td
(TcRPA-1)Tj
3.9458 0 Td
(and)Tj
1.7292 0 Td
(TcRPA-2\))Tj
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(and)Tj
1.7348 0 Td
(showed)Tj
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(that)Tj
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(this)Tj
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(protein)Tj
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(complex)Tj
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(is)Tj
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(indeed)Tj
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(a)Tj
0.652 0 Td
(single)Tj
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(stranded)Tj
3.7134 0 Td
(DNA)Tj
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(binding)Tj
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(protein)Tj
3.1521 0 Td
(that)Tj
1.7801 0 Td
(interacts)Tj
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(with)Tj
1.9956 0 Td
(DNA)Tj
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(through)Tj
3.4412 0 Td
(the)Tj
1.4684 0 Td
(TcRPA-1)Tj
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(subunit.)Tj
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(Then,)Tj
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(we)Tj
1.3266 0 Td
(suggest)Tj
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(that)Tj
1.7745 0 Td
(TcRPA)Tj
3.1181 0 Td
(participates)Tj
4.8302 0 Td
(in)Tj
1.0318 0 Td
(canonical)Tj
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(functions)Tj
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(in)Tj
1.0375 0 Td
(DNA)Tj
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(metabolism,)Tj
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(such)Tj
2.0579 0 Td
(as)Tj
1.0148 0 Td
(replication)Tj
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(and)Tj
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(repair)Tj
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(pathways.)Tj
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(we)Tj
1.3266 0 Td
(were)Tj
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(able)Tj
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(to)Tj
1.0261 0 Td
(show)Tj
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(that)Tj
1.7802 0 Td
(the)Tj
1.4683 0 Td
(reduction)Tj
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(of)Tj
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(TcRPA-2)Tj
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(expression)Tj
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(by)Tj
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(heterozygous)Tj
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(knockout)Tj
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(generation)Tj
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(slowed)Tj
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(down)Tj
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(S-phase)Tj
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(and)Tj
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(conse-)Tj
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(quently)Tj
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(cell)Tj
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(proliferation.)Tj
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4.3937 0 Td
(while)Tj
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(cell)Tj
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(growth)Tj
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(was)Tj
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(impaired)Tj
3.8494 0 Td
(in)Tj
1.0318 0 Td
(TcRPA-2)Tj
3.9458 0 Td
(heterozygous)Tj
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(knockout)Tj
4.0251 0 Td
(cells,)Tj
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(a)Tj
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(better)Tj
2.5399 0 Td
(efficiency)Tj
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(in)Tj
1.0375 0 Td
(differentiation)Tj
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(from)Tj
2.211 0 Td
(epimastigote)Tj
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(to)Tj
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(metacyclic)Tj
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(trypomas-)Tj
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(tigote)Tj
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(forms)Tj
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(and)Tj
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(metacyclic)Tj
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(trypomastigote)Tj
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(infection)Tj
3.7928 0 Td
(was)Tj
1.7007 0 Td
(observed,)Tj
4.0195 0 Td
(suggesting)Tj
4.405 0 Td
(that)Tj
1.7745 0 Td
(the)Tj
1.474 0 Td
(delay)Tj
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(of)Tj
1.0147 0 Td
(cell)Tj
1.5591 0 Td
(cycle)Tj
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(progression)Tj
4.9209 0 Td
(is)Tj
0.8504 0 Td
(linked)Tj
2.7269 0 Td
(to)Tj
1.0262 0 Td
(differentiation)Tj
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(in)Tj
/F14 1 Tf
1.0318 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
/F14 1 Tf
1.0375 0 Td
[(\n)1()-9(\r)]TJ
/F9 1 Tf
(.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 222.0094 cm
BT
/F3 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Methods)Tj
/F0 1 Tf
0 -1.5023 Td
(Modeling)Tj
4.2189 0 Td
(and)Tj
1.8661 0 Td
(molecular)Tj
4.4977 0 Td
(dynamics)Tj
4.3985 0 Td
(simulations)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 186.9732 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(A)Tj
0.9014 0 Td
(flexible)Tj
3.0897 0 Td
(linker)Tj
2.5738 0 Td
(containing)Tj
4.5241 0 Td
(21)Tj
1.1735 0 Td
(residues)Tj
3.481 0 Td
(that)Tj
1.7801 0 Td
(connect)Tj
3.3902 0 Td
(OBF2)Tj
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(and)Tj
1.7291 0 Td
(OBF3)Tj
2.5512 0 Td
(on)Tj
1.2756 0 Td
(TcRPA-1)Tj
3.9458 0 Td
(and)Tj
1.7291 0 Td
(its)Tj
1.1452 0 Td
(20)Tj
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(C-terminal)Tj
4.6431 0 Td
(residues)Tj
3.4809 0 Td
(do)Tj
1.2529 0 Td
(not)Tj
1.5761 0 Td
(show)Tj
2.3017 0 Td
(homology)Tj
4.2519 0 Td
(with)Tj
1.9956 0 Td
(any)Tj
1.6611 0 Td
(protein)Tj
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(DNA-binding)Tj
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(structure.)Tj
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(fore,)Tj
2.0352 0 Td
(we)Tj
1.3266 0 Td
(created)Tj
3.1238 0 Td
(separated)Tj
4.0081 0 Td
(models)Tj
3.1068 0 Td
(for)Tj
1.3833 0 Td
(OBF1/OBF2)Tj
5.21 0 Td
(of)Tj
1.0148 0 Td
(TcRPA-1)Tj
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(\(TcRPA-1-OBF12)Tj
7.4438 0 Td
(model,)Tj
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(corresponding)Tj
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(to)Tj
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(the)Tj
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(1254)Tj
2.6362 0 Td
(region\))Tj
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(and)Tj
1.7291 0 Td
(for)Tj
1.3833 0 Td
(OBF3)Tj
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(of)Tj
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(TcRPA-1)Tj
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(\(TcRPA-1-OBF3)Tj
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(model,)Tj
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(corre-)Tj
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(sponding)Tj
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(to)Tj
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(the)Tj
1.4683 0 Td
(272445)Tj
3.5887 0 Td
(region\).)Tj
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(A)Tj
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(of)Tj
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(the)Tj
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(crystal)Tj
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(structure)Tj
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(of)Tj
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(DBD-A)Tj
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(and)Tj
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(DBD-B)Tj
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(of)Tj
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(RPA1)Tj
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(from)Tj
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2.2053 0 Td
[()31()11( )6()]TJ
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[()-3()12()11(\r)4()2()]TJ
/F9 1 Tf
3.0274 0 Td
(\(PDB)Tj
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(ID)Tj
1.2983 0 Td
(1JMC;)Tj
2.8063 0 Td
([)Tj
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(20)Tj
0 g
(]\))Tj
2.1883 0 Td
(was)Tj
1.7008 0 Td
(selected)Tj
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(as)Tj
1.0148 0 Td
(the)Tj
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(best)Tj
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(template)Tj
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(for)Tj
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(an)Tj
1.2018 0 Td
(initial)Tj
/F14 1 Tf
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[(\r)12()]TJ
1.0147 0 Td
[()10(\r)4()12(\r)4(\n)7()]TJ
/F9 1 Tf
2.1544 0 Td
(model)Tj
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(for)Tj
1.389 0 Td
(TcRPA-1-OBF12)Tj
7.0979 0 Td
(\(score:)Tj
2.8743 0 Td
(342.7;)Tj
2.5738 0 Td
(E-value:)Tj
3.4526 0 Td
(3e-57;)Tj
2.6419 0 Td
(identity:)Tj
3.5319 0 Td
(41%\).)Tj
2.4945 0 Td
(Chain)Tj
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(C)Tj
-34.6278 -1.3039 Td
(\(Replication)Tj
5.1136 0 Td
(protein)Tj
3.1521 0 Td
(A)Tj
0.9071 0 Td
(32)Tj
1.1679 0 Td
(kDa)Tj
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(subunit;)Tj
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(RPA2\))Tj
2.9026 0 Td
(of)Tj
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(the)Tj
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(crystal)Tj
2.8119 0 Td
(structure)Tj
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(of)Tj
1.0148 0 Td
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