Chapter 16 THE MOLECULAR BASIS OF INHERITANCE

GENES CARRY INFORMATION

Early geneticists thought that genes were made of proteins.
Archibald Garrod (1908) introduced the idea that genes and enzymes are related.
iscussed the genetic disease al!aptoneuria.
"omogentisic acid ("#)$ an intermediate of the %rea!down of phenylalanine and
tyrosine$ is e&creted in the urine.
'arrod theorized that an enzyme that o&idizes "# was lac!ing and that this was due to a
mutation of the gene.
James Sm!er (19(6) showed that enzymes were proteins.
Frederic" Gri##i$h (19(8) con)erted a)irulent pneumococcus to the )irulent strain.
E&periment with mice in*ected with a)irulent li)e cells and heat+!illed )irulent cells.
#)irulent cells were con)erted to )irulent cells.
Tra!s#orma$io! due to a transforming principle
O%T% A&er'( C%M% McLeod and M% McCar$' (19,,) identified the transforming principle to %e
-#.
Geor)e Beadle and Ed*ard Ta$m (19,0s) suggested that a single gene specifies each
protein.
.hey wor!ed with fungus -eurospora crassa.
-eurospora is a haploid organism.
/ne gene$ one protein hypothesis.
Al#red Hershe' and Mar$ha Chase (190() conducted e&periments on the reproduction of
%acteriophages.
.hey showed that -# enters the cell.
-# is re1uired for synthesis of new protein coats and -#.
Er*i! Char)a## (1900) determined the composition of -#2 ratios of adenine+thymine and
guanine+cytosine were )ery close to 1.
Rosali!d Fra!"li! and M%H% +il"i!s conducted e&periments on the &+ray diffraction of the -#
molecule in the early 003s.
James +a$so! and Fra!cis Cric" (1904) proposed a model for the structure of the -#
molecule %ased on the wor! done %y 5ran!lin and 6il!ins. .hey proposed the doble heli,
structure.
-EO.YRIBONUCLEIC ACI- / -NA
1. -#$ a polymer$ is made of two polynucleotide chains intertwined to form a doble heli,.
(. Each nucleotide monomer contains a nitrogenous %ase which may %e one of the
0ri!es1 ade!i!e or )a!i!e
or
0'rimidi!e1 $h'mi!e or c'$osi!e.
4. Each %ase is co)alently lin!ed to deo&yri%ose$ a 0C sugar.
,. eo&yri%ose is co)alently %onded to a phosphate.
0. .he %ac!%one of each single -# chain is formed %y alternating deo&yri%ose and
phosphate groups *oined %y 2hos2hodies$er lin!ages.
6. Each phosphate group is lin!ed to the 03 car%on of one deo&yri%ose and to the 43 car%on of
the other deo&yri%ose.
7. "ydrogen %onds form %etween adenine and thymine (two %onds)$ and %etween guanine and
cytosine (three %onds). .he se1uence of %ases is complementary %ut not identical. .his
allows to predict the se1uence %ases in one strand if one !nows the se1uence of %ases in
the other strand.
8. Each pair %ase is 0.4, nm from the ad*acent pair %ases.
9. .here are ten %ase pairs in each turn of the heli& ma!ing each turn 4., nm high.
10. .he dou%le heli& is ( nm wide.
11. .he chains run in an opposite direction and are said to %e a!$i2arallel to each other. #t the
end of each -# molecule there is an e&posed 03 car%on on one strand and an e&posed 43
car%on on the other strand.
1(. Complementary %ase paring of adenine and thymine and guanine and cytosine are the
%asis of Chargaff 8s rule$ which is # 9 . and C 9 . in -#.
-NA RE0LICATION
# process called re2lica$io! can precisely copy -#.
.he essential features of -# replication are uni)ersal %ut there are some differences %etween
pro!aryotes and eu!aryotes due to the difference in -# organization.
:n pro!aryotes$ -# consists of a circular dou%le+stranded molecule$ while in eu!aryotes it is
made of a linear dou%le+stranded molecule associated with great deal of proteins.
.he two strands of the dou%le heli& unwind. Each strands ser)es as a template for the formation
of a new complementary strand.
-# replication is semico!ser&a$i&e2 each daughter dou%le heli& contains one strand from the
parent -# and one newly synthesized strand.
;ore than a dozen enzymes and proteins are in)ol)ed in -# replication.
MECHANISM
1. -# %egins at specific sites in the molecule named ori)i!s o# re2lica$io! and forms the
replication %u%%le. "ere at each end of the replication %u%%le$ -# helicase creates a
re2lica$io! #or".
(. .he position of the replication for! is constantly mo)ing as replication proceeds.
4. .he enzyme -NA helicase tra)els along the heli& opening it as they mo)e.
,. Si!)le/s$ra!d bi!di!) 2ro$ei!s %ind to the single -# strands pre)enting reformation of
the dou%le heli&.
0. To2oisomerases %rea! and re*oin sections of the -# to relie)e strain and pre)ent !nots
during replication.
6. -# synthesis always proceeds in a 03 43 direction2 0< phosphate at one end and 4<
hydro&yl at another end.
7. .he two -# strands are a!$i2arallel$ that is$ their sugar phosphate %ac!%ones run in
opposite directions.
8. -NA 2ol'merases catalyze the lin!ing together of the nucleotide su%units. .here are at
least ele)en -# polymerases in)ol)ed in eu!aryote replication.
9. -ucleotides with three phosphate groups are used as su%strates for the polymerization
reaction. .wo of the phosphates are remo)ed and the nucleotide is added to the 43 end of
the growing strand.
10. .hese reactions are e&ergonic and do not re1uire #.=.
11. -# polymerase cannot initiate the synthesis of polynucleotide> they can only add
nucleotides to the 43 end of an already e&isting chain that is %ase+paired with the template
strand.
1(. -# synthesis re1uires an RNA 2rimer to initiate the synthesis reaction. .he ?-# primer is
made of a%out ten nucleotide long in eu!aryotes.
14. .he ?-# primer is synthesized %y a protein comple& !nown as a 2rimosome$ which
includes an enzyme$ 2rimase$ that is a%le to start a new strand of -# opposite a -#
strand.
1,. -# replication is continuous in one strand and discontinuous in the other.
10. -# polymerase adds nucleotides to the 43 of the new strand that is always )ro*i!)
$o*ard the replication for!. .his strand is called the leadi!) s$ra!d%
16. -# polymerase adds nucleotides to the 43 of the new strand that is )ro*i!) a*a' from
the replication for!. .his strand is called the la))i!) s$ra!d. .he rate of elongation is the
addition of a%out 000 nucleotides per second in %acteria and 00 in humans.
17. =rimase synthesizes a short ?-# primer$ which is e&tended %y -# polymerase to form an
O"a3a"i #ra)me!$.
18. .he lagging strand is synthesized in short pieces called O"a3a"i #ra)me!$s$ which are
made of 100 to 1000 nucleotides. .he fragments were disco)ered %y ?ei*ii /!aza!i.
19. .hese fragments grow in a direction away from the replication for!.
(0. Each /!aza!i fragment %egins with an ?-# primer.
(1. #fter it has %een elongated %y -# polymerase$ the ?-# primer is degraded$ the gaps are
filled with -# and the ad*oining fragments are lin!ed together %y -NA li)ase.
((. -# ligase lin!s the 43 end of one fragment with the 03 end of the ad*oining fragment.
-# replication is bidirec$io!al starting at the origin of replication and proceeding in %oth
directions.
#n eu!aryotic chromosome may ha)e se)eral origins of replication and may %e replicating at
se)eral points at any one time.
EN4YMES RE0AIRS ERRORS
-# polymerase proofreads each nucleotide against its template as soon as it is added. :f there
is an error$ the nucleotide is remo)ed and the correct one is added in its place.
Errors that arise after replication are also corrected.
Ncleo$ide e,cisio! re2air.
.he mismatch pair of nucleotide distorts the -# molecule>
# !clease enzyme cuts the damaged -# strand at two points>
.he -# is repaired %y -NA 2ol'merase %y filling the gap with the correct nucleotides>
-NA li)ase attaches the new se1uence to the rest of the molecule.
EU5ARYOTIC CHROMOSOMES
?eplication results in the formation of a chromosome with two dou%le helices.
Each dou%le heli& corresponds to a chromatid.
Eu!aryotic chromosomal -# molecules ha)e special nucleotide se1uences called telomeres
at their ends.
.hese end caps of repetiti)e -# are called $elomeres.
.elomeres do not contain genes. .hey are made of multiple repetitions of a nucleotide
se1uence$ e. g. ..#''' is the repetiti)e unit in humans.
?ecent research supports the idea that the re2e$i$i&e -NA at the end of the chromosome has a
protecti)e function.
.he num%er or repetitions in a telomere )aries from 100 to 1000.
# small amount of telomeric -# fails to replicate each time the -# replicates. -o essential
genetic information is lost.
.elomeric -# can %e lengthened %y a -# replicating enzyme called $elomerase%
.elomerase molecules ha)e a small ?-# molecule together with the protein.
Cells that produce telomerase continue to di)ide indefinitely %eyond the point at which cell
di)ision would normally cease.
#cti)e telomerase is found in germ cells that gi)e rise to sperm and eggs in animals$ %ut it is
a%sent in somatic cells.
.he a%sence of telomerase acti)ity in animal cells may %e the cause of cellular aging.
:t is possi%le that telomeres are a limiting factor in the life span of certain tissues.
A CHROMOSOME CONSISTS OF -NA AN- 0ROTEINS
Chromatin consists of -@ and histones. Chromatin is 10 nm thic!.
=roteins called his$o!es are responsi%le for the first le)el of -# pac!ing.
;ost of the histone amino acids are positi)ely charged (lysine or arginine) and %ind tightly to the
negati)ely charge -#.
-# winds twice around the histones and form a !cleosome. -ucleosomes resem%le %eads
in a string. .he -# %etween nucleosomes is called the li!"er.
.he amino end of each histone e&tends outward from the nucleosome. .his is the -+terminus.
.he histone tails of one nucleosome and the lin!er -# and nucleosomes on either side
interact and cause the -# to coil forming a chromatin fi%er 40 nm thic!.
.he 40+nm fi%er$ in turn$ forms loops called loo2 domai!s. Aoop domains are 400 nm thic!.
.he loops are attached to a -# scaffold made of proteins.
:n a mitotic chromosome$ the loop domains coil and fold further compacting the chromatin to
produce the mitotic -# found in the metaphase of cell di)ision.
;etaphase chromosomes are a%out 700 nm thic!.
@B;;#?C
Cou must !now the function of the following enzymes2
-# helicase
@ingle+strand %inding proteins
.opoisomerases
-# polymerases
=rimase
-uclease
Aigase
.elomerase
#lso the structure$ function and meaning of the following2
=rimer 43 end of chromosome (phosphate)
Aeading strand 03 end of chromosome (hydro&yl)
Aagging strand antiparallel
/!aza!i fragment %idirectional
.elomeres replicating for!
?epetiti)e -# semiconser)ati)e
Chromatin replication
"istones
-ucleosomes
Ain!er
Aoop domain