Nucleic acids as genetic information carriers

DNA as the Source of Genetic Information

Although DNA was discovered in 1869 b !riedrich "iescher as a new# acidic# $hos$horus containing substance made u$ of ver large molecules that he named %nuclein&# its biological role was not recogni'ed( In 1889 )ichard Altmann introduced the term %nucleic acid&( * 19++ the $urine and $ rimidine bases were ,nown( -went ears later# the two ,inds of nucleic acids# )NA and DNA# were distinguished( An incidental but $recise observation .19/80 and relevant investigations .19110 indicated that DNA could be the carrier of genetic information(

Discovery of transformation
-he idea that genetic material is nucleic acid had its roots in the discover of transformation in 19/8( -he $henomenon was first observed in 19/8 b !red Griffith# an 2nglish $h sician whose s$ecial interest was the bacterium that causes $neumonia# Streptococcus pneumonia.

Transformation
Griffith had succeeded in isolating several different strains of S. pneumonia .t $e I# II# III# and so forth0( In the virulent .disease causing0 forms of a strain# each bacterium is surrounded b a $ol saccharide coat# which ma,es the bacterial colon a$$ear smooth when grown on an agar $late3 these forms are referred to as S# for smooth( Griffith found that these virulent forms occasionall mutated to non4virulent forms# which lac, a $ol saccharide coat and $roduce a rough a$$earing colon on an agar $late3 these forms are referred to as )# for rough(

Transformation
Griffith finall concluded that the t $e II) bacteria had somehow been transformed, ac5uiring the genetic virulence of the dead t $e IIIS bacteria( -his transformation had $roduced a $ermanent# genetic change in the bacteria3 though Griffith didn6t understand the nature of transformation# he theori'ed that some substance in the $ol saccharide coat of the dead bacteria might be res$onsible( 7e called this substance the transforming principle.

Identification of the transforming $rinci$le

After 1+ ears of research# 8swald Aver # 9olin "ac:eod# and "acl n "c9art succeeded in isolating and $urif ing the transforming substance( .19110( -heir a$$roach to the $roblem was to chemicall destro all the ma;or categories of chemicals in the e<tract of dead cells one at a time and find out if the e<tract had lost the abilit to transform( -he virulent cells had a smooth $ol saccharide coat# whereas the nonvirulent cells did not3 hence# $ol saccharides were an obvious candidate for the transforming agent( 7owever# when $ol saccharides were destro ed# the mi<ture could still transform(

-ransforming =rinci$le
=roteins# fats# and ribonucleic acids .)NA0 were all similarl shown not to be the transforming agent( -he mi<ture lost its transforming abilit onl when the donor mi<ture was treated with the en' me deo< ribonuclease .DNase0# which brea,s u$ DNA( -hese results strongl im$licate DNA as the genetic material( 7owever# man biologists still refused to acce$t the idea# however# still $referring the h $othesis that the genetic material is $rotein(

-he 7ershe 49hase 2<$eriment

A second $iece of evidence im$licating DNA as the genetic material resulted during the stud of the bacterium Escherichia coli and one of its infecting viruses bacterio$hage -/ conducted b Alfred 7ershe and "artha 9hase( -/ is a bacteriophage .$hage0 that infects the bacterium Escherichia coli.

-he 7ershe 49hase 2<$eriment

At the time of the 7ershe 49hase stud .their $a$er was $ublished in 19>/0# biologists did not understand e<actl how $hages re$roduce( ?hat the did ,now was that the -/ $hage consists of a$$ro<imatel >+@ $rotein and >+@ nucleic acid# that a $hage infects a cell b first attaching to the cell wall# and that $rogen $hages are ultimatel $roduced within the cell( *ecause the $rogen carried the same traits as the infecting $hage# genetic material from the infecting $hage must be transmitted to the $rogen # but how this occurs was un,nown

-he 7ershe 49hase 2<$eriment

7ershe and 9hase designed a series of e<$eriments to determine whether the $hage protein or the $hage DNA was transmitted in $hage re$roduction( -o follow the fate of $rotein and DNA# the used radioactive forms (isotopes) of $hos$horus .A/=0 and sulfur .A>S0( -he labeled the ca$sular $rotein of bacterio$hages with radioactive sulfur .A>S0 and the DNA with radioactive $hos$horus .A/=0( ?hen bacteria were infected with the labeled bacterio$hage# onl A/= .DNA0 entered the cells# and not the A>S .ca$sular $rotein0( -he subse5uent formation of new# com$lete $hage $articles in the cell $roved that DNA was the e<clusive carrier of the genetic information needed to form new $hage $articles# including their ca$sular $rotein(

-ransfection 2<$eriments
In 19>B# several re$orts demonstrated that if 2( coli was treated with the en' me l so' me( -he outer wall of the cell can be removed wvithout destro ing the bacterium( 2n' maticall treatcd cells are na,ed so to s$ea,( and contain onl the cell mcmbrane as their outer boundar ( Such structures arc called $roto$lasts .or s$hero$lasts0( Cohn S$ililen and Dean !raser inde$endentl rc$orted that b using s$hero$lasts the were able to initiate $hage re$roduction with disru$ted -/ $articles(

-ransfection 2<$eriments
Similar( but more refined# e<$eriments were re$orted in 196+ b George Guthrie and )obert Sinsheimer( DNA was $urified from bacterio$hage .DE 1B1( a small $hage that contains a single stranded circular DNA molecule of some >A86 nucleotides( ?hen added to 2( coli $roto$lasts the $urified DNA resulted in the $roduction of com$lete DE 1B1 bacterio$hagcs( -his $rocess of infection b onl the viral nucleic acid( called transfection $roved conclusivel that DE 1B1 DNA alone contains all the necessar information for $roduction of mature virus(

)NA As Genetic "aterial

A few viruses utili'e )NA# not DNA# as their genetic material( -his fact was demonstrated in 19>6 b 7ein' !raen,el49onrat and *ea Singer# who wor,ed with tobacco mosaic virus .-"F0# a virus that infects and causes disease in tobacco $lants( -"F $ossesses a single molecule of )NA surrounded b a helicall arranged c linder of $rotein molecules( !raen,al49onrat found that# after se$arating the )NA and $rotein of -"F# he could remi< them and obtain intact# infectious viral $articles

)NA As Genetic "aterial

In their wor,# )NA and coat $roteins were se$arated and isolated from -"F and a second viral strain# 7olmes ribgrass .7)0( -hen# mi<ed viruses were reconstituted from the )NA of one strain and the $rotein of the other( ?hen this Gh bridG virus was s$read on tobacco leaves# the lesions that develo$ed corres$onded to the t $e of )NA in the reconstituted virus4that is( viruses with wild4t $e -"F )NA and 7) $rotein coats $roduced -"F lesions and vice versa( Again# it was concluded that )NA serves as the genetic material in these viruses(

)NA As Genetic "aterial

Also in 19>6# Alfred Gierer and Gerhard Schramm demonstrated that )NA isolated from -"F is sufficient to infect tobacco $lants and direct the $roduction of new -"F $articles# confirming that )NA carries genetic instructions(