THE BEGINNINGS OF MOLECULAR GENETICS: GENE FUNCTION

Discuss from a historical perspective the discoveries that led to the hypothesis that one gene defines one polypeptide List and describe several genetic defects leading to enzyme deficiencies in humans Explain how genes determine the structure of proteins and the effects of mutations that alter a proteins structure

GENE CONTROL OF ENZYME STRUCTURE

Garrods Hypothesis of Inborn Errors of Metabolism One Gene-One Enzyme Hypothesis

Alkaptonuria
Phenylketonuria Albinism Lesch-Nyhan Syndrome Tay-Sachs Disease

GENE CONTROL OF PROTEIN STRUCTURE

Sickle-Cell Anemia Cystic Fibrosis

Garrods Hypothesis of Inborn Errors of Metabolism

English physician Archibald Garrod Provided the first evidence (1902) that genes were related to enzymes with his studies on patients with alkaptonuria From his observations, Garrod concluded that alkaptonuria is a genetic disease caused by the absence of a particular enzyme necessary for the metabolism of homogentisic acid Since the mutation was recessive, only homozygotes express the defect

One Gene-One Enzyme Hypothesis

George Beadle and Edward Tatum (1942) showed in their studies with the fungus Neurospora crassa that there was a direct relationship between genes and enzymes Beginning of Biochemical Genetics They proposed the one gene-one enzyme hypothesis, which states that a specific gene controls the production of a single active enzyme

Life cycle of the haploid, mycelialform fungus Neurospora crassa

Method devised by Beadle and Tatum to isolate auxotrophic mutations in Neurospora

Methionine biosynthetic pathway showing four genes in Neurospora crassa that code for the enzymes that catalyze each reaction

One Gene-One Enzyme Hypothesis

Mutations that result in the loss of the enzyme activity lead to the accumulation of precursors in a metabolic pathway and an absence of end product Enzymes are proteins that can consist of more than one polypeptide, and genes encode individual polypeptide chains The modern description of this hypothesis then is: one gene-one polypeptide

Phenylalanine-tyrosine metabolic pathways

Inborn errors of metabolism in breakdown of phenylalanine and tyrosine

Phenylketonuria

Phenylketonuria (PKU) is caused by an autosomal recessive mutation in the gene encoding the enzyme

phenylalanine hydroxylase that occurs in about 1 of every 12,000 Caucasian births

Phenylketonuria

Without

this enzyme, phenylalanine (an essential amino acid) is converted to phenylpyruvic acid, which affects cells of the central nervous system causing mental retardation, slow growth, and early death

person with PKU can live normally if phenylalanine is restricted from the diet

Albinism

Albinism is caused by an autosomal recessive mutation in a gene coding an enzyme (tyrosinase) that converts tyrosine to the brown pigment melanin
1:33,000 Caucasians- 1:28,000 African American

Without melanin, albinos have white skin and hair, and red eyes They are more sensitive to light because melanin plays a role in protecting the skin against UV light from the sun

An albino monkey

Red eyes and white skin and hair in an albino

Albinism

At

least two different mutations are known that occur in different steps of the melanin pathway if two albinos have children, the children can be normal if the mutations complement

Thus

Lesch-Nyhan Syndrome

Lesch-Nyhan Syndrome is caused by a recessive mutation in the gene encoding hypoxanthine guanine phosphoribosyl transferase (HGPRT), a gene on the X chromosome
1:10,000 males are affected

HGPRT is involved in a purine utilization pathway

Lesch-Nyhan Syndrome

When it is absent, excess purine bases

accumulate and are converted to uric acid which builds up in the body

Feeding

disorder, mental retardation and self-mutilation are characteristic of this disease, and it results in premature death (before the age of 20)

A case of mental retardation due to Lesch-Nyhan Syndrome

Tay-Sachs Disease

Tay-Sachs Disease is a lysosomalstorage disease that is caused by a recessive mutation in the gene encoding the enzyme N-acetylhexoaminidase A It is rare in the population at large but high in Ashkenazi Jews N-acetylhexoaminidase A cleaves a terminal N-acetylhexoaminidase group of a brain ganglioside

Tay-Sachs Disease

Without

the enzyme, unprocessed gangliosides accumulate in brain cells, leading to cerebral degeneration and death by age 3 is rapid neurological degeneration associated with this disease that leads to generalized paralysis, blindness, loss of hearing, and premature death

There

The biochemical step for the conversion of the brain ganglioside GM2 to the ganglioside GM3, catalyzed by the enzyme N-acetylhexosaminidase A (hex A)

A child with Tay-Sachs

Accumulation of gangliosides in the brain cells

GENE CONTROL OF PROTEIN STRUCTURE

Sickle-Cell Anemia: Symptoms and Causes

Sickle-cell anemia is the result of a mutation in a gene encoding the polypeptide subunit of hemoglobin In the polypeptide chain, the amino acid glutamic acid is replaced by valine, a substitution that is caused by a point mutation in the gene

The hemoglobin molecule

Sickle-Cell Anemia: Symptoms and Causes

This autosomal co-dominant mutation causes a single amino acid substitution in the chain which alters the chemical structure of hemoglobin inside of red blood cells and changes the oxygen carrying capabilities of the molecule Red blood cells carrying the mutant hemoglobin are sickle-cell shaped, more fragile than normal cells and not as flexible

The first seven N-terminal amino acids in normal and sickled hemoglobin polypeptides

Electrophoresis

of hemoglobin
variants

Other Hemoglobin Mutants

Over two-hundred hemoglobin mutants

in the or chains have been
detected, and several are well

characterized

Examples of amino acid substitutions found in polypeptides of various human hemoglobin variants

Examples of amino acid substitutions found in polypeptides of various human hemoglobin variants

Cystic Fibrosis

Cystic Fibrosis (CF) is caused by an autosomal recessive mutation in a gene that encodes the CF protein 1:2,000 Caucasians, 1:23 heterozygous carrier (most common lethal autosomal recessive) 1:90,000 Asians The defective gene was initially identified by RFLP mapping on chromosome 7 rather than biochemical testing Normal Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a chloride channel in some types of cell membranes

Proposed structure for cystic fibrosis transmembrane conductance regulator (CFTR)

Cystic Fibrosis

In people with CF, the mutated gene encodes an abnormal CFTR protein, which causes abnormal salt transport across membranes This leads to the accumulation of mucus in the lungs and pancreas and other symptoms Lethal disease: Life expectancy about 40 yrs of age

THE END