Page 1 \ ?^p 6r.1 CELL CYCLE CONTROL OF HUMAN H4 ...
Page 1 \ ?^p 6r.1 CELL CYCLE CONTROL OF HUMAN H4 ...
Page 1 \ ?^p 6r.1 CELL CYCLE CONTROL OF HUMAN H4 ...
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Mt F: Cett C\.1e Catud cn Ha Ger. I6nsciotion<br />
O/CDP 1 and HjNF P/<strong>H4</strong>-TF2 thal bind ro the Sile ll cell cycle domain.<br />
Howe!€r, th€ contriburions ol lh€se proroios to rhe mdimal level ol <strong>H4</strong> gene<br />
transcriplion during rhe c€ll cycl€ remains io b€ €stabilshed. In the slldies<br />
d€scribed in rhis chapter, a seloded ser of mutanr <strong>H4</strong> promoref reporte.g€n€<br />
constucls which were staby lnregrated into the genome of hlman HsLa s3<br />
c€lls were used, and analyzed N4 prornot€r aclvity during lhe cetcyce n<br />
synchronized cells. In contast lo prevlous suggesuons, oL/ cellcycls snalysis<br />
reveals rhal mutarion ol rho H|NF.M/|RF.2 binding sire (mutant Msp-16) which<br />
dramatically reduces rhe rat6 ot historc g6m iranscripiion, does not atrer ths<br />
cerl cycle @nvolled enhancemont ol <strong>H4</strong> gene promole. aclivily during earty S<br />
phase. MLta{on ol the HiNF,D/CDP-1 binding sile (murant SUB-j1) or<br />
combi.€d muHion oi all thro€ hislone gene ranscription faclor bindi.g siles<br />
{mutanl l\,lPM-17) has subtle bLrt slatislca ty signifcant etiecrs on either tho<br />
timng or lhe magnirude of <strong>H4</strong> g€ne transcfiplion rale. ]t is conctud€ that<br />
lemporal conrroi ot <strong>H4</strong> g6n€ ifanscription dlring the celt cycte apparenly<br />
rsquires the integrated activilles ot mult ple ranscription factors.<br />
Ulilization ol RNase proiecrlon assry3 to. studying <strong>H4</strong><br />
gen€ lransc.iprion durrng the celt cycte<br />
IrasciipUonal control ol lh6 <strong>H4</strong> gene ranscription has previously b6€6<br />
analyzed using run,on rranscriprion anatyses (Ramsey-Ewing er a/., 1994).<br />
However, it was obsetoed that ihis rechniqus undsr the current erperim€ntal<br />
condrions does nor represed a r€li€bt€ m€rhod tor moniloring subr6<br />
vanarons in <strong>H4</strong> gene rransc.iplon This s due in pad to ab€tranly targe<br />
lluclLrations in radiogreph c signals to..ibosomatRNA synlrests which is used<br />
140
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