Page 1 \ ?^p 6r.1 CELL CYCLE CONTROL OF HUMAN H4 ...

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A2t2 F Cer Ctrte cannd d tlr Gde r@apt'a ransq pllonal conirol during lhe @ll cycl€- lh6 liEt obleciiv6 ol this study was lo assBss rhe i.dividla contributions ol Hlstone Nuclear Factors (HiNF's) interacling wilh <strong>H4</strong>-site ll ro the level ol transc ion. S€condlv we sludLed involvem€nr ot these HiNFs in $e l€mporal .egulation ol histone gene rranscriplion d! ng th€ cellcycle. Whil€ lhis work was in operalion a.umber ol oth6r labo€lories hav6 also ini at€d and/or codlnued sludios on histon€ g€m otganizaton and expresson, For examPle, s€v€ral hlsion6 g6n€s wars recenlly clon€d and charact€rizod in many ditlerBnl sp€cies (Albig d a/ , 1991 ; Oobn€r er a/.' 1991 i Drab€nt et a/, 1991, 1993r Nakayama €.d S€roguchi,, 1991i Oonq er €/- 1994). l. addton, lho €rPrBssion pattsrns ol disrinct hislone geres has been analyzod d! ng diflgr€.lalion (Koppel €l a/., 1994i lvanova er 3/ l994ai Budeind s? at., rs4i albig el a/., 1993i Col'ad er a/., 1992) or i6 ths c€ll cycle (Famsey el a, ., 19o5; Brocard el a/., 1994r H€ints. 1991i wolle and GriFes, 1993; choi and chas., 1993i Kremer and Kis er, 1992i Eilers ea a/' 1994i sreinel€/. 1994ivanden E el a/., 1994; Fams€y el a/ , 1994; Boute.ia d a/., 1993j gonner er a/., 1993; Lim and cha€., 19924 Gom€z-coadrado €t a/ 1992). Fudhefinor€, a nLrmb€r ol cis-acling 6lemer't hsve been described io' many hisrone qenes (Hatch and aonn6r., 1995; lvanova er 3/ 1994b1 Khochbin and t?wrence, 1994; Ohshig€ el a/, 1993r Khochbin and wolre, 1993t Hlnkley and Perry, 1992i G mes€la/,19S2: ChOi and chae 1991; Wolto and Grlmes, 1991; van winon €13/, 199la b,c) and m€ cognate vans' acti.g iactors have also been lde.lili€d (Madinelli and H€inlz 1994i van w',nenerar 1904. Sh€@ood el a,., 1993, van W',qnen el a/ 1992abc 1991i 27
A.E F: Cet CEl. Cotuot ot <strong>H4</strong> t3.M f@btbn Huh er at, 1991 I Palb €l a/,, 1S). D$pil6 lhb $densivo s€l of dala oblai.ed by difierent hboralodes sp6daliznE on hlslorE genl3, hduding lh€ stein labo.atory, the work pr€s6nt6d ln lhis th€sb rspresants th6 frrst 6vld6nc6 tof rhe imponanco ot rhs co.nbin€d adn ni€s ol rruhbl€ lactoG in the rcgulation ot hisrone gene 6xprosion d!.ing rhe c€! q/d., a
Page 1 and 2: \ ?^p 6r.1 CELL CYCLE CONTROL OF HU
Page 3 and 4: In tho narn€ of Alah, olt G6cloua
Page 5 and 6: Acknowl6dg6msnts Contents Abstract
Page 7 and 8: Selection ol cell linss contalning
Page 9 and 10: ACKNOWLEDGEMENTS
Page 11 and 12: I would also lkg ro rhank Dr. Pafic
Page 13 and 14: ABSTRACT
Page 15 and 16: i.leraclion elem€nt, sil6 ll, Thr
Page 17 and 18: 8. Colony hybridizalion sc.eening o
Page 19 and 20: INTRODUCTION
Page 21 and 22: t t F: con cda c@tot d H. G@ r@Mbn
Page 23 and 24: Azh F: C6n Ca/cro C@t.t ot H. G.n.
Page 25 and 26: Aziz f: Celi Ctcle Cotutol ol H4 Ae
Page 27 and 28: = 2. q t6l tl9t E L:I la-lf titl I.
Page 29 and 30: En F. Ce Cycl. Cantd d h. Gft ra.sc
Page 31 and 32: ,r1z F Cett cy.te connot ol H4 Gene
Page 33 and 34: A2i2F: Cettcdt cotubtot e@IrNiotid
Page 35 and 36: Ntz F: Ce Cwte Contd d 114 Gm tansc
Page 37 and 38: t- s- i;> f z x + n q e o, { T T z
Page 39 and 40: AzzF cett Crct. ContolatH4Gen ldn*t
Page 41 and 42: a2i2 F cdt c$t. connd d H4 Gm Tdnsc
Page 43 and 44: o rl$orrE GEIE FEEI{IXU =; FrN -!r
Page 45: !e 3lhllrllon c.I cycl. control aa
Page 49 and 50: Crct C@nd d H. GNf,'/MAi@ '2izE:Ce
Page 51 and 52: 9C| -= = , o:li?eiP :< :::? il il 7
Page 53 and 54: s.tudi. dhcrm or do"rn! -l /4 I I I
Page 55 and 56: aziz F: cel crd. cotuot d 81 cd. fn
Page 57 and 58: a2lz F: celt cyd. c.ntol d H4 G@. f
Page 59 and 60: A2iz F Cett cycta contat al Hl Gene
Page 61 and 62: E2 F. cen crct. Connat al H4 c*e Ta
Page 63 and 64: Colony hybridization 4a aa a a
Page 65 and 66: ,dd f . c.t cfi|e cotud d H4 cd'e f
Page 67 and 68: Restriction Analysis Of Mutants e'b
Page 69 and 70: ,d, F: ccl crc!. cqfrol ot H10d fdt
Page 71 and 72: AtIzF: Cd CJ.L Cdtuotol BaGq:€lat
Page 73 and 74: ,.8 F: 6t W @tlol o, rk 0.,t rn',df
Page 75 and 76: ldD F: Cett CrL Cotuot at t14 G* Iw
Page 77 and 78: A2E F. cdt crcto Cantat at Ha Gqa r
Page 79 and 80: Aziz F: Cdl Ced. Cd*ol ol H4 Gse ra
Page 81 and 82: o E o !!tH I rtog 5l €s c_g - t-
Page 83 and 84: kir F: cettctttd contal at H4 cen.
Page 85 and 86: arE F: CeI Crct Cotuatat 114 G40 fa
Page 87 and 88: c A B e'l' t i _ f '', \J..$.'
Page 89 and 90: Nz F: Cel Ctd. Ctuol at h4 Cq. faBd
Page 91 and 92: Aziz F: Celt Cyct. Contd d H4 6@e l
Page 93 and 94: azE F. celt cy.b contol al H4 Gene
Page 95 and 96: A Geneticin (G4'18) rssistanl HeLa
Page 97 and 98:
NizF: Cen qcb cttuat al tu G@ ft|tl
Page 99 and 100:
M F: Cd W W.l h. 4.. 7'.t'll'Pn'd F
Page 101 and 102:
'dit F: ce qcle cotuot ol Ha Gm ra@
Page 103 and 104:
M. F: Ce|| Crd. Cqnrot ot Ha Gerc T
Page 105 and 106:
azi. F: cetl cttcL ctu.t or Ha Gtu
Page 107 and 108:
kiz F: c.lt
Page 109 and 110:
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
Page 115 and 116:
High mitotic index alter relsase tr
Page 117 and 118:
ND F Cen atct6 cannot ot B4 6ene rr
Page 119 and 120:
NE F: Cell C6t cotuat .t H4 cffi fh
Page 121 and 122:
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 and 174:
E Comparison ol TgT9 ratios lor cAT
Page 175 and 176:
Azd F: Cd Ctcle Ca\ttol ot ltt Gtuf
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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