Studies on Nucleic Acids in Relation to Protein Synthesis

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( 1 ) Several methods f o r t h c f r ? . c t i o n e t i @ n of t r a n s f e r IWA have been i n v e s t i g a t e d . These i n c l u d e c o u n t e r c u r r e n t d i s t r i b u t i o n , i o n

exchange chromatogra.phy, chemiccl methods u t i l i z i n g She; ox!- d e t i o n of t ~ z n s f e r RNA. w i t h p e r i o d a t e, and h y d r o ~ e n bonr'ing method-

.

( 2 ) T'le e f f e c t o f t z q x x r e t u r e on t h e i o n exckange f r a c t i o n a t i o n c h e r a c t e r i ; t i c s of y e a s t t r a n s f e r RITA hag been studied using t h e a n i o n exchanger d i e t h y l a m i n o e t h y l - c e l l u l o s e (DUE--cellulose).

(

z*)

Â ^methodôf p r e p a r e t i v e f r a c t i o n a t i o n of transfer _{RNA has}been d e v e l o p e d , T r a n s f e r RNA was e l u t e d from DEAE-cellulose columns a t 55' ûsinga g r e d i e n t of i n c r e a s i n g s o d i u m c h l o r i d e . The f r a c t i o n - e t e d RIGA 3howcd c o n s i d e r a b l e p u r i f i c a t i o n when individual f r a c t i o n s were as8ayc.d f o r amino _{a c i d}acceptance u s i n g an enzyme preparation d e r i v e d from e i t h e r yezst c e l l s o r f r o m EF&cherichia

-

c o l i cells.

I ~ P ~ l t i p l e peeks o f a c c e p t o r a c t i v i t y f o r t h e sane amino a c i d were o b s e m e d for s e v e r a l a m i n o a c i d s ,

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^A second metbo2 of p r e p a m t i v e f r e c t i o n q t i o n of t r a n s f e r XNA is d e s c r i b e d using DEAE-Seplxdex The t r z z s f er _BBTA was e l u t e d from t 3 e column with a b u f f e r o f c o n s t a n t sodium chloride

c o n c e n t r e t i o n and a decreasing t e m p e r a t u r e g r a d i e n t from 65 0 to

5 5 ' .

The o r d e r o f e l u t i o l ? o f d i f f e r e n t amino a c i d a c c e p t i n g transfer RnA s p e c i e s was found t o d i f f e r f r o m t h a t obtained by DmGcellulose chromatography,

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^By ^{u s e}^of ^t?le^above^methods^{f o r}f r a c t i u n a t i m in c o c j u r i c t i o n w i t h c o u n t e r - c u r r e n t d i a t r i b u t i o r ~ a c c o r d i n g t o t h c method of

Z a c h m

& g .

^(1961),highly p u r i f i ~ d y e a s t t r c n s f e r 81,rA spcciees a c c ~ p t i n g g l y c i r x , s e r i n c , v a l i n e , l y s i n e and p h e n y l a l a n i n e were o b t a i n e d . Thc m i r n i m u m p u r i t i e s o f t h e s e t r a n s f e r RNA s p e c i e s were f o u n d t o h~ 61$, 447$

51$,

817b and

7860,

r ~ s p e c t i v c l y .

( 6 ) ^RNA i s o l a t ~ d f r o m b o t h b a k e r s ' cnc brewers

'

^{y ~ s t}^was f r a c t i o n - a t e d using DEIiE-Scphadex. B u l t i p l e p m k s o f v a l i n e a c c e p t a n c e

a c t i v i t y wcrc dctectci? u s i n g enzyme r,repar;ztions d c r i v c r ? from bakcrs

'

y ~ c s t , hrcwsrs4 y ~ a s t and EP c c l i . The r c s u l t ~ a r c d i s - cussed i n tcms o f t k s p e c i f i c i t y of t h c zlmino a c y l RNA s s y n t h ~ t a s e s f o r t r a n s f ~ r R%A0

( 7 ) E n z y m p r e p a r a t i o n s werc n ~ d e f r o n

5

c o l i i n f c c t e d w i t h b a c t e r i o 2 h a g e Tq and from n o n - i n f e c t e d

--

E. c o l i . Thc ~ b i l i t y

of cech p r ~ p a r a t i o n t o a n i n o a c y l a i e EL c o l i t r a n s f e r RFA was c o m p r c d f o r z number of amino zicidso TLE r e s u l t s 2re d i s c u s s e d in r c l a t i n n t o tha c f f ' e c t o f T 4 ?hage infection on p r o t e i n s y n t h e s i s in

--

E. coli.

( 8 ) ^The s t i n u l s t i o n o f p r o t c i n s y n t h e s i s by polyuridylic a c i d and by t u r n i p y e l l o w mosaic v i r u s liNA wcre compared in a c e l l free p r o t e i n s y n t h e s i s systei? d e r i v d from E. c o l i .

( 9 )

The hybridization p r o p e r t i e s o f

-- E.

c o l i transfer _and ribosomal RITA vwcrc examined u s i n g d c m t u r ~ d E. c o l i DNA i r f i o b i l i z e d on

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1 ' '

cellulose n i t r z t c f i l t c r s . r h c two nhA c l r s n c s h y l r i d i z c d

c o n p c t i t i v c l y f o r s i t c 3 on i h a Dh'I

.

^The p o c s i b l c s i g n i f i c a n c e o f t h i s r ~ s u l t for t h c c o n t r o l of s y n t l ~ e s i s of t r m s f c r and

r i b o s o m d RLA i s ciiscussc5,

( 1 0 ) The i n c o r p o x t i o n of r e d - i o a s t i v c p r e c u r s o r s i n t o

--

^E.

--

^{c o l i}

t r ~ i : i s f c r iU% VRS s t u d i c d by fr?.,ctioi-Lnting r t i d i c ~ c t i v e

--

E. c o l i RDA or. D E h E c d l u l o s e c o l m s n a t 65'. Chcnses i n i n c o r p o r a t i o n wcsc d c t c c t e c f n l i o x i ~ l g i c d u c t i o n of the c;nr,;-r,e R - f ; c l ~ c t o s i d a s e , 2nd i n f c c t i c n by b e . c t e r i e p h a g r . s IlS22 and 1'4.

(11) Lovr c o l e c u l a r m i g h t T d pha; c s p ~ c i f i c RKA ms prcnent i n TA plmgc i n f c c t e d

&

c o l i . K study of its o r o p c r t i c s l e d to t h e c o n c l u s i o n s t h r t i t ncs different t o TL -?hag9 mssscngcr RSA end had EEVJ of t h f p r o p c x t i e s ~ x p c c t e d of trcnsf sr FCNA. The

2ossible origin'= of t h i s l o w molrculzr w e i g h t R L ~ a r c discussed in terms o f r r _ o - s ~ n ~ ; c . r NL.A metabolism a n d its controlo

( 1 2 ) A s h o r t r c v i c w cf t h c p r o c e s s e s of c o n t r o l of genc e x p r e s s i o n is p r c s c - c i c d ,

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