By N. B. Myant

Ldl cholesterol Metabolism, LDL, and the LDL Receptor

summary: ldl cholesterol Metabolism, LDL, and the LDL Receptor

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Extra info for Cholesterol Metabolism, Ldl, and the Ldl Receptor

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1978 ) hav e als o show ntha t heptane-extracte dLD Lparticle s reconstitutedwit h25-hydroxycholesteryl oleat e (25HC-LDL) ar e abou tfou rtime sa seffectiv ea s LD Lreconstitute dwit hcho ­ lesteryl oleat e (C-LDL )i nsuppressin g reductas e activit yi nculture dnorma l humanfibroblasts whe nequa l quantitieso f 25-hydroxycholesterolan dcholester ­ ol ar e internalize db y th e cells . Whe nnorma lfibroblast s ar e grow ni nth e pres ­ enceo f 25HC-LD Lwit hn o externa l sourc eo f cholesterol , the y ceas e growin g anddi e withi na fe w days .

Osborneet al. (1987 ) hav e show ntha tsterol-regulate dexpressio no f th e hamsterreductas e gen e als o requires th e bindin go f transcriptionfactor s t o spe cific sequence si nth e promoterregion . Usin g th e metho do f DNas eI footprinting (see Chapte r9 , Sectio nIV) , the yidentifie dsi x region s (FP 1 t o FP6 )i nth e 227 bppromote rtha twer e protecte dfro mDNas eI digestio nb y th e bindin go f proteins presen ti nhamster-livernucle i (Fig . 13) . Th e rol eo f thes e region si n transcriptiono f th e reductas e gen e wa s investigate db y constructin g plasmid s containing reductase-CATfusio ngene si nwhic hth e nucleotid e sequence si nth e protectedregion s wer e altere db y oligonucleotide-directedmutagenesi s (se e Sec tionV, Ebelow) .

T h e multipl e transcriptio n initia tionsite s predicte d b yS1 nucleas e analysi s ar e indicate d b y n u m b e r e d bar s a b o v e th e n u c l e o t i d e ; thos e predicte d b y prime r extensio n ar e indicate d b y dot s b e l o w th e nucleotide . Repeated sequence s C C G C C C ar e enclose d b y boxes ; th e sequenc e G G G C G Gi s s h o w n b y lines a b o v e an d b e l o w th e sequence . T h e position s o f eigh t A T G codon s 5 ' t o th e initiato r A T G c o d o n ar e indicate d b y M e t b e l o w th e sequence .

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