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aj| F. Cdt Cttt. C@ud ot H1 cetu rnnsu,p!,@ It was furrher €xamined rhat lhe signficance ol reductions in rc/rg ratos lor the CAT rnFNAsignats obse.ved with 1wo ot the mutantH4 promotsr constucls. Full scale cel cycte synchfonjes with tour difierenr c6I linss wef6 perlomed (Figs. 3740) lo address whether reduclions in lhe redued T3/m fanos ar€ r€Lated to changes in the maqniiude ot hision€ gene irascipnon during ea y s phase, or wherher this ratio r€lects a shft in lhe timing oi rh6 peak ol H4 prcnoter aclivity to tor exampta eartier or tster me-points, The RNas6 prot€crion assays show that CAT mRNA tevets diven by r're I!,tSp-16 mltantH4 promor€r,which can not bind HiNF-M/|FF2. ar€ vory simitar to CAT mnNAlevels obssrved lor lho witdryp€ H4 promoter FO1O8 celline (Figs.37 and 38). In com€st, mutant suBrl which is not capabt€ ot bindtng HiNF-D, appears t0 display a modesl delay in the riming of maamat H4 promoier adlvity (Fig. 38). Fu.rhermore, rhe muiani MprMrT consruq appears ro navs a smilar limlnq in lhe peak ol rnaxlrnat tansc.ipton, but dspays pantaL retention ol cell qde dependent etevations in H4 gene ransc.iplton (Fig,4O), ll is concluded that cell cycle conl.ol represents a comptex mechanism which nvolr'es s-bne co_Lbu0ons oimurripl€di nct pror€,ns 1!i
Azd F: Cd Ctcle Ca\ttol ot ltt Gtufan*npti@ Flg.37. R€r"h cell cyct6 rnalysls ot thewltd tlyps H4 promoter cAf stabte cslt llns. (A) t3Hl Thymidins incorporation was m€asured at th€ lndi€at€d Imes lollowing fsleas6 lrom sscord thynidine uock as d6scrib6d in Maleials and Merhods B€sults ars €xp'€ssed as clunrs per minuls ot acid prcdPitat€d pn1 t'vmloi* 1e) RNass proteclion alsay ol H4 CAT and GAPDH RNA as described in Figw€s 30'32 (C) Ouantitatlon of th€ assays in pad A ushg a Belascop€ analvz6r and Prssented as perc€nl maximum oata points a'6 the mean valu€s trom M0 independent expedments and th€ ba6 repcsent the differences betu€en each pan ol r€spectivs values The signal associated with caPDH mBNA did not aler signilicantly throughoul lne 156
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\ ?^p 6r.1 CELL CYCLE CONTROL OF HU
Page 3 and 4:
In tho narn€ of Alah, olt G6cloua
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Acknowl6dg6msnts Contents Abstract
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Selection ol cell linss contalning
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ACKNOWLEDGEMENTS
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I would also lkg ro rhank Dr. Pafic
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ABSTRACT
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i.leraclion elem€nt, sil6 ll, Thr
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8. Colony hybridizalion sc.eening o
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INTRODUCTION
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t t F: con cda c@tot d H. G@ r@Mbn
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Azh F: C6n Ca/cro C@t.t ot H. G.n.
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Aziz f: Celi Ctcle Cotutol ol H4 Ae
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= 2. q t6l tl9t E L:I la-lf titl I.
Page 29 and 30:
En F. Ce Cycl. Cantd d h. Gft ra.sc
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,r1z F Cett cy.te connot ol H4 Gene
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A2i2F: Cettcdt cotubtot e@IrNiotid
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Ntz F: Ce Cwte Contd d 114 Gm tansc
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t- s- i;> f z x + n q e o, { T T z
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AzzF cett Crct. ContolatH4Gen ldn*t
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a2i2 F cdt c$t. connd d H4 Gm Tdnsc
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o rl$orrE GEIE FEEI{IXU =; FrN -!r
Page 45 and 46:
!e 3lhllrllon c.I cycl. control aa
Page 47 and 48:
A.E F: Cet CEl. Cotuot ot H4 t3.M f
Page 49 and 50:
Crct C@nd d H. GNf,'/MAi@ '2izE:Ce
Page 51 and 52:
9C| -= = , o:li?eiP :< :::? il il 7
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s.tudi. dhcrm or do"rn! -l /4 I I I
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aziz F: cel crd. cotuot d 81 cd. fn
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a2lz F: celt cyd. c.ntol d H4 G@. f
Page 59 and 60:
A2iz F Cett cycta contat al Hl Gene
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E2 F. cen crct. Connat al H4 c*e Ta
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Colony hybridization 4a aa a a
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,dd f . c.t cfi|e cotud d H4 cd'e f
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Restriction Analysis Of Mutants e'b
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,d, F: ccl crc!. cqfrol ot H10d fdt
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AtIzF: Cd CJ.L Cdtuotol BaGq:€lat
Page 73 and 74:
,.8 F: 6t W @tlol o, rk 0.,t rn',df
Page 75 and 76:
ldD F: Cett CrL Cotuot at t14 G* Iw
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A2E F. cdt crcto Cantat at Ha Gqa r
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Aziz F: Cdl Ced. Cd*ol ol H4 Gse ra
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o E o !!tH I rtog 5l €s c_g - t-
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kir F: cettctttd contal at H4 cen.
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arE F: CeI Crct Cotuatat 114 G40 fa
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c A B e'l' t i _ f '', \J..$.'
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Nz F: Cel Ctd. Ctuol at h4 Cq. faBd
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Aziz F: Celt Cyct. Contd d H4 6@e l
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azE F. celt cy.b contol al H4 Gene
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A Geneticin (G4'18) rssistanl HeLa
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NizF: Cen qcb cttuat al tu G@ ft|tl
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M F: Cd W W.l h. 4.. 7'.t'll'Pn'd F
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'dit F: ce qcle cotuot ol Ha Gm ra@
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M. F: Ce|| Crd. Cqnrot ot Ha Gerc T
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azi. F: cetl cttcL ctu.t or Ha Gtu
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kiz F: c.lt
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ktz F. cen ctct6 ctuat at tu 6e rft
Page 111 and 112:
Azi2 F: Cen Cvcte Contot .t H4 Gere
Page 113 and 114:
I 200000 1mmo DNA Synth.3l3 In Sync
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High mitotic index alter relsase tr
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ND F Cen atct6 cannot ot B4 6ene rr
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NE F: Cell C6t cotuat .t H4 cffi fh
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A22 F: Cett cytl. Connd d H4 64. rn
Page 123 and 124: hnF cet cyttd cannot ot B4 6end ltu
Page 125 and 126: Nuclear Run-on TranscriPlion wr Fol
Page 127 and 128: NlzF Cett cr.t. canuat oI H4 Gene f
Page 129 and 130: RNA isolated f rom cell synchrony H
Page 131 and 132: A2iz F: c.u cwL contat al Ha GeN f@
Page 133 and 134: RESULTS
Page 135 and 136: trb F: cett clcL cotuot al H4 Gen.
Page 137 and 138: M2F:Co Crrt6 Cdrtot ot H4G.n. fn.pi
Page 139 and 140: HiNF-tvl HiNF-P HiNF-D random DNA p
Page 141 and 142: andom DNA poly G/C competitor oligo
Page 143 and 144: a2i2 F: ceu ctcte contol al H4 Gene
Page 145 and 146: aziz F: c.ll crd. cdtd 0t H4 Gene t
Page 147 and 148: compeutor oligonucleolides: Site ll
Page 149 and 150: A1i2 F: Ceu a:rcte cotuot ot H1 G@
Page 151 and 152: andom DNA poly G/C competilor oligo
Page 153 and 154: EEF:Ce Olcta Cont t ol114 Gene l@ns
Page 155 and 156: , F: cal qcb cotfrtl d rh 9.n ftuaf
Page 157 and 158: kL E d N. Cdfr.t ol k e.rE ltlr&tu
Page 159 and 160: Mt F: Cett C\.1e Catud cn Ha Ger. I
Page 161 and 162: A.*F: Cen q.h cdtuotot Ha 6an ranfl
Page 163 and 164: 3!j{99!caaw arir c",. r_*rooo FI9 3
Page 165 and 166: I -a2b F: cel cyde c@xol d B1 aa@ h
Page 167 and 168: -= GAPDH mRNA = = 012 66 hours afle
Page 169 and 170: Fq *' I E t li! lil tlr ilt Frlr li
Page 171 and 172: (! (! t- (Y) F o 'F (E d z l t52 +
Page 173: E Comparison ol TgT9 ratios lor cAT
Page 177 and 178: :tE &I qiE{, ccaoi oftaa Cer tir fi
Page 179 and 180: kEE@q.l.co''o,a,tl,od,',rutg/bt!,.
Page 181 and 182: ibEc|,Otd.c!,'t,dlHao'tllep,rcl Fg.
Page 183 and 184: DrscussroN
Page 185 and 186: I azz F Con CYb Cnror ot H4 G@e ntn
Page 187 and 188: ,@2 F. Cdt CEle Cotud .l U1 G.N fan
Page 189 and 190: l NE P C.l qE/€ Cotud d H4 Gw f@f
Page 191 and 192: aznF:couct l. cdtold H16@f@tioti@ s
Page 193 and 194: ari2 F: c.t] cyctd contnt ot H1 Ger
Page 195 and 196: Atiz F. Cdl cycl6co.ttol d H4 Gene
Page 197 and 198: l A2izF C.ttctct (onntt ot a. calo
Page 199 and 200: azi. F: cei c\cb cotuot d H4 G@ fon
Page 201 and 202: NizF Can a$t. c@tot ot H4 G.n fftNc
Page 203 and 204: Atiz F: Catl Cvcte contot ol H4 6.n
Page 205 and 206: Aztz F a.tt ctcte conr ot .t Hl 6en
Page 207 and 208: ,rit 9: Ceu Cycle C.htot ot Ha C.n6
Page 209 and 210: tai. F: ceu cycte cantat ot ,r1 Gen
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