BIOL0003 Lecture 05: The Chromosome Theory and Sex Linkage

I will let you into a secret; the genes are on the chromosomes. Now, of course

– have complete sequence of many chromosomes; this seems a truism. But –

how do we know? In fact, classic idea of what I think of as “genetical thinking”:

inference of deep biological truth by clever thinking rather than brute force!

Mendel - no concern with physical nature of the gene: simply a particle, with no

information as to where in the cell it might reside. However, an immediate

question: where are the genes? Chromosome theory surprisingly contentious for

a long time. Largely because chromosomes visible for only a small part of the

cell cycle (will not discuss in detail here: assume everyone has heard of mitosis

and meiosis).

Chromosomes cannot be seen in interphase and might, it was, supposed, simply

be dissolved. Also, even in the early history of genetics, became obvious that

there are far more genes than there are chromosomes – can they be the same

thing? What is evidence that genes are held on chromosomes, and what is

implication of this?

General Correspondence Between Genes and Chromosomes

Early in C20 - chromosome behaviour established. Immediate parallel with

Mendel’s “particles”: Just like genes chromosomes:

1) Present in pairs (we have two copies of chr 1, chr 23 etc).

fertilisation.

3) Different chromosomes segregate independently.

Also, some plants with extra chromosomes show heritable differences in

appearance, suggesting that genes were associated with chromosomes. All very

interesting, but scarcely a proof. However, the proof soon arrived.

Sex Linkage and the Proof of the Chromosome Theory.

To Mendel made no difference which is the male and which the female parent:

pollen from green with egg from yellow pea gives same result as pollen from

yellow with egg from green.

However, Thomas Hunt Morgan, fly lab, Columbia University, around 1916.

Some reciprocal crosses in Drosophila gave very different results. Mutant turned

up in one stock: changed eye colour from red to white.

Crosses gave very different result, depending on which parent was red and

which white.

1) Red female x White male - F1 All red

2) Red male x white female - F1 All females red, all males white!
Mate the F1 flies together; another odd result.

Cross 1) F2 All females red, half males red and half white.

Cross 2) F2 Half females red, half females white, half males red, half males

white.

At first sight, baffling. But - male and female Drosophila differ in one

chromosome pair, the sex chromosomes. Discovered Nettie Stevens 1905

Females - two large X chromosomes, XX (ie all eggs are X).

Males - a large X and a much smaller Y, XY (ie half sperm are X and half are Y).

Hypothesis: the locus for eye colour is carried on the X. Females have two alleles

at this locus, males only one.

Cross 1) would then be:

Male Male

w
X Y X Y
w
X XX XY X XX XY
Female w --------> w w w
X XX XY X XX X Y

Cross 2) would be

Male Male
 w
X Y YX
w w w w
X XX X Y X XX XY
Female w w W --------> w w w w
X XX X Y X XX XY

In other words, the pattern of inheritance of the white allele - until then a

hypothetical structure - follows exactly that of the X chromosome, a physical

object. Strong evidence that genes are indeed borne on chromosomes. Now -

two sorts of inheritance; sex-linked, as above, and autosomal - on non-sex

chromosomes.

Soon, a definite proof. Morgan and one of his students noticed that, in some

crosses, the above pattern did not apply. Nearly always, white females with red

males give all daughters red and all sons white in the F1, as in cross 2.

Bafflingly, though, in a few cases, there was the opposite results: white females

mated with red males gave all daughters WHITE, all sons RED! Morgan:

“Treasure your exceptions!”. Examined stock: found that their chromosomes

were abnormal. The female parents are ANEUPLOIDS - because of an error

(nondisjunction, failure of the X chromosomes to separate) during cell division

they produce eggs which are XX or have no sex chromosomes at all (Normal

females of course just produce eggs with one X). The cross is then:

XY
w w ww ww
XX XXX XXY
DIES WHITE FEMALE

O XO YO
 RED DIES
MALE

Ie a chromosomal abnormality has led to a matching genetic abnormality.

[Should note that the sex-determination mechanism is different in Drosophila and

humans: XO and XXX humans are viable and female, XXY are viable and male.

This is because in insects it is the number of Xs that determine sex: 2 = female, 1

= male. In mammals, though, it is the presence of a Y chromosome (that carries

a male-determining gene) that is essential. Ie XXY or even XXXXXXXXY is a

male, XO a female. The two systems are less similar than they seem].

The final confirmation: a white-eyed stock arose in which the females have their

two X chromosomes stuck together, so they are inherited as a unit (the

ATTACHED-X stock). They ALWAYS gave this anomalous result: that is, when

crossed to red males, they gave white daughters and red sons. Their attached X

chromosomes are transmitted as one unit, and so too are their two copies of the

white allele. A clear proof that the gene for white eye (whose existence has been

inferred only from breeding experiments) is congruent with structures visible

down the microscope - chromosomes.

Remarkably enough, birds and butterflies have the opposite pattern of

chromosomal sex determination. Males are the homogametic (XX, in fact

referred to as WW for clarity) sex, females the heterogametic (XY, referred to as

Drosophila: again, a proof of the chromosome theory.

Now know many characters in both Drosophila and humans that show X-linked

inheritance. A very few even show Y-linkage.

Sex-linked Inheritance

Recessive alleles on the X: characteristic pattern - transmitted mainly through

females, manifest mainly in males. This is because males need only one copy of

the allele (they are hemizygous) to show its effects; females need two.

Character usually much more common in male.

Also, for affected male, none of his sons are affected, but all his daughters are

carriers. Means that half his grandsons are affected. None of his sons pass on

the gene, as they carry his Y chromosome, not his X.

Many examples known in humans; has a characteristic pattern: Haemophilia:

Blood clotting disorder (in fact several different kinds). Blood in joints,

extravasated blood. Famous case - descendants of Queen Victoria: her 8th child,

Leopold, affected with the disease. Two of her daughters must have been

carriers as they had affected male descendants. Best-known family: that of

Tsarina Alexandra, QV’s grand-daughter; married to Tsar Nicholas II of Russia;

son, Tsar Alexis severely affected. Others - got into Spanish royal family through

Victoria Eugenie, Ena. Ironic, as she was chosen to infuse new blood into the
Bourbon line: Philip V, unbalanced, sensual; Ferdinand IV - mad and impotent;

Ironic that harmful gene brought in from the mother’s side of the family.

Notable that no history of the disease in British Royal Family before Victoria

(although one of her mediaeval ancestors known, for reasons obscure, as

Ludovicus the Skinless). Claim by clergyman that haemophilia caused by use of

chloroform in childbirth, as “robs God of the deep earnest cries which arise in

times of trouble for help”. Now know, though, that it was due to mutation - almost

certainly in the august testicles of Edward, Duke of Kent, her father. Will return

to this later.

Certainly not in present British royal family, as descends through male line from

Victoria. By now probably lost from Victoria’s female descendants, too: no

haemophiliac sons born for >60 years.

Now - treatment with factor VIII cures symptoms; notable - this another eg where

a genetic disease is treated with an environmental change.

2) Lesch-Nyhan syndrome: disease of purine metabolism; low IQ and self-

mutilation.

3) Red-green colourblindness; cannot separate red from three

3) Sex-linked dominants - rare; but one is a type of vitamin D-resistant rickets.

Will leave to you to work out pattern of inheritance.

One obvious problem with X-linkage: if males manage with one copy of a gene,

how do females cope with two (or, to put it the other way, if females have two,

how can males manage with only one?). We know that usually an extra

chromosome is very harmful and a missing one is lethal. Why does not one sex

die because of chromosomal imbalance?

A dosage compensation mechanism - an introduction to the idea that genes can

be regulated - switched on and off. Very important in study of development (see

later). X chromosome inactivation (XCI), or Lyonisation, after Mary Lyon, who

discovered it.

In female mammals, early in development, one of the Xs in a particular cell is

switched off: visible as a dark-staining blob in every cell (the Barr Body).

The number of Barr Bodies is always one less than the number of X

chromosomes. Means that every adult female is a mosaic: a mixture of cells,

some with one and some with the other X chromosome active. If she is a

heterozygote, different patches of cell have one or the other allele expressed.

Every patch is a clone descending from an early embryonic cell, with one or other
of the Xs inactivated. Often refer to Xm - the X of maternal origin - and Xp, the X

of paternal origin.

Familiar example - tortoise-shell cat: X linked locus O (orange or black alleles, O

and o), Patches of orange and black hair on skin.

Humans – sex-linked red-green colourblindess.

Females heterozygous for red-green colourblindness: have effectively normal

vision (can separate red from green). However, if sweep laser across retina,

switching from red to green, and asking colour, there are patches of cells that are

colour blind, and patches that are not. Ie mosaic nature of retina. Often quite

different patterns of expression in left eye and right eye.

Most sex-linked diseases; limited to male. Well-known example: Duchenne

muscular dystrophy. Wasting disease of muscles; mutations in the dystrophin

gene - enormous. Very few female cases indeed, as would have to be

homozygotes (one copy of the gene from mother, one from father - but DMD

boys die before sexual maturity).

However, as in tortoiseshell cat, female heterozygotes may activate some of the

dystrophy genes in their own cells – usually this has no discernible effect; ie in

effect the MD allele is recessive. However, in some cases, the inactivation takes
place early in development, and many of her cells may have the MD allele active,

with some features of the disease (ie in effect a case of incomplete dominance.

One remarkable case - two girl identical twins, one with DMD, one without.

Turned out that both were heterozygotes, but in one, early in development, the X

with the DMD gene on it had been activated, reaching many of her muscle cells,

while in the other, the alternative X, with the normal allele, was active.

XCI under control of a single gene, which forms an “inactivation centre”,

switching off genes near it. In fact, those a long way away may not be fully

inactivated. Recently found locus - the Xist locus; X inactivation specific

transcript – large (17kb) RNA binds on to inactivated X. Under control of an X

inactivation centre on the ACTIVE X. As far as we can see, which one is activa

and which inactive is random.

Notable that Drosophila does the same thing with sex-linked genes to ensure

equality of action in males and females but in a totally different way. Instead of

randomly inactivating half the X chromosomes, all X chromosomes are switched

on, but those of males are set at an activity twice that of females. Quite a

different gene involved - sex-lethal: different alleles kill either males or females,

depending on whether they over-activate or under-activate the genes on the X.

how different it is (in spite of apparent similarities) in humans and Drosophila.

Others - even more bizarre; eg alligators, sex determined by developmental

temperature, fish by social pressure.

Nevertheless, sex-determination and the X chromosome gave the fundamental

insight into the location of the genes, on chromosomes. Will pursue this in the

next several lectures.

HUMAN CHROMOSOMES

CELL CYCLE

MITOSIS

GENERAL CORRESPONDENCE OF GENES AND CHROMOSOMES

MORGAN’S LAB

DROSOPHILA MALE AND FEMALE

NETTIE STEVENS AND THE Y CHROMOSOME

RED AND WHITE EYE

MORGAN’S CROSS

MORGAN CONTINUED INTO

MORGAN FIRST CROSS INTERPRETED

MORGAN SECOND DITTO

MORGAN’S ANOMALOUS RESULT STATED

MORGAN’S ANOMALOUS RESULT ANALYSED

X CHROMOSOME

CLOTTING CASCADE

FACTOR VIII

HAEMOPHILIA – LEAKAGE

VICTORIA AND ALBERT

VICTORIA FAMILY

HAEMOPHILIA PEDIGREE
CLOSE UP ON RUSSIAN BRANCH

ALEXIS

RASPUTIN

LESCH NYHAN

ISHIHARA TEST DEMONSTRATION

X LINKED DOMINANT PEDIGREE

VISIONS OF RED AND GREEN

ISHIHARA TEST DEMONSTRATION

ISHIHARA TEST

RESULT REVEALED

BARR BODY

MARY LYON

XIST – X INACTIVATION SPECIFIC TRANSCRIPT RNA

TORTOISESHELL FEMALE AND ORANGE BROTHER

CONES IN EYE

XCI IN COLOURBLIND WOMAN

LEFT AND RIGHT EYES

MUSCULAR DYSTROPHY MUSCLES

DYSTROPHIN

MD SYMPTOMS

MD AND NORMAL MUSCLE CELLS

HET FEMALE SHOWING XCI