Chapter 3 The Chromosomal Basis of Heredity Chromosomes • The chromosome complement = the complete set of chromosomes of plants and animals • The nucleus of each somatic cell contains a fixed number of chromosomes typical of the particular species • The number of chromosomes vary tremendously among species and have little relationship to the complexity of the organism 2 Chromosomes • The chromosomes in the nuclei of somatic cells are usually present in pairs. For example, the 46 chromosomes of human being consist of 23 pairs • Cells with nuclei of this sort, containing two similar sets of chromosomes, are called diploid 3 Chromosomes • The germ cells, or gametes, are haploid and contain only one set of chromosomes, consisting of one member of each of the pairs • The haploid gametes unite in fertilization to produce the diploid state of somatic cell • The chromosomes are present in pairs because one chromosome of each pair derives from the maternal parent of the organism and the other from its paternal parent 4 Mitosis • Mitosis is a precise process of nuclear division that ensures that each of two daughter cells receives a diploid complement of chromosomes identical with the diploid complement of the parent cell • Mitosis is usually accompanied by cytokinesis, the process in which the cell itself divides to yield two daughter cells 5 Cell Cycle • In a cell that is not undergoing mitosis, the chromosomes are invisible with a light microscope. This stage of the cell cycle is called interphase • DNA in the chromosomes is replicated during a period of interphase called S = DNA synthesis • Before and after S, there are periods, called G1 and G2, respectively • These three interphase periods are followed by mitosis, M 6 Figure 3.2: The cell cycle of a typical mammalian cell growing in tissue culture 7 Stages of Mitosis • Prophase is marked by the condensation of chromosomes. Each chromosome is already longitudinally double, consisting of two subunits called chromatids • Each pair of chromatids is the product of the duplication of one chromosome in the S period of interphase • The chromatids in a pair are held together at a specific region of the chromosome called the centromere. 8 Prophase of Haemanthus © Andrew S. Bajer - Research Projects 9 Stages of Mitosis • At the beginning of metaphase, the mitotic spindle forms • The spindle is a bipolar structure arching between the centrosomes that consists of microtubules • The spindle fibers attach to each chromosome in the region of the centromere at the kinetochore • The chromosomes move toward the center of the cell until all the kinetochores lie on an imaginary plane equidistant from the spindle poles = the metaphase plate 10 Metaphase of Haemanthus © Andrew S. Bajer - Research Projects 11 Stages of Mitosis • In anaphase, the centromeres divide longitudinally, and the two sister chromatids of each chromosome move toward opposite poles of the spindle • Once the centromeres divide, each sister chromatid is regarded as a separate chromosome in its own right. 12 Anaphase of Haemanthus © Andrew S. Bajer - Research Projects 13 Stages of Mitosis • In telophase, a nuclear envelope forms around each compact group of chromosomes, nucleoli are formed, and the spindle disappears • The chromosomes undergo decondensation until they are no longer visible as discrete entities • The two daughter nuclei assume a typical interphase appearance • The cytoplasm of the cell divides in two 14 Telophase of Haemanthus © Andrew S. Bajer - Research Projects 15 Figure 3.3: Chromosome behavior during mitosis in an organism with two 16 pairs of chromosomes Meiosis • Meiosis is a mode of cell division in which cells are created that contain only one member of each pair of chromosomes • Meiosis consists of two successive nuclear divisions • Meiosis results in four daughter cells, each genetically different and each containing one haploid set of chromosomes • Meiosis is a more complex and considerably longer process than mitosis and usually requires days or even weeks 17 Meiosis • In animals, meiosis takes place in specific cells called meiocytes • The oocytes form egg cells and the spermatocytes form sperm cells • In the females of both animals and plants, only one of the four products develops into a functional cell (the other three disintegrate) 18 Figure 3.5: The life cycle of a typical animal 19 Meiosis • In plants, the products of meiosis form spores, which undergo one or more mitotic divisions to produce a haploid gametophyte organism • The gametophyte produces gametes by mitotic division of a haploid nucleus • Fusion of haploid gametes creates a diploid zygote that develops into the sporophyte plant, which undergoes meiosis to produce spores and so restarts the cycle 20 Figure 3.6: The life cycle of corn, Zea mays 21 Outline of Meiosis • Prior to the first nuclear division, the members of each pair of chromosomes become closely associated along their length • The chromosomes that pair with each other are said to be homologous chromosomes • Each member of a pair of homologs consists of a duplex of two sister chromatids joined at the centromere. The pairing of the homologous chromosomes, therefore, produces a fourstranded structure 22 Outline of Meiosis • At the time of pairing, the homologs can exchange genes that results in chromosomes that consist of segments from one homolog intermixed with segments from the other • In the first nuclear division, the homologous chromosomes are separated from each other, one member of each pair going to opposite poles of the spindle • Two nuclei are formed, each containing a haploid set of duplex chromosomes 23 Outline of Meiosis • The second nuclear division resembles a mitotic division, but there is no DNA replication • At metaphase, the chromosomes align on the metaphase plate, and at anaphase, the chromatids are separated into opposite daughter nuclei • The net effect of the two divisions is the creation of four haploid nuclei, each containing the equivalent of a single sister chromatid from each pair of homologous chromosomes Figure 3.4: Behavior of a single pair of homologous chromosomes in meiosis 24 Mitosis vs. Meiosis • Meiosis produces four cells: each contains one copy of each pair of homologous chromosomes = genetically different, haploid • Mitosis produces two cells that contain both members of each pair of homologous chromosomes = genetically identical, diploid 25 Meiosis I • The first meiotic division—reductional division, reduces the chromosome number by half • Prophase I is the longest stage and is commonly divided into five substages: leptotene, zygotene, pachytene, diplotene, and diakinesis These are descriptive terms that indicate the appearance of the chromosomes at each substage 26 Meiosis: Prophase I • Leptotene – the chromosomes first become visible as long, thread-like structures Figure 3.8A: Leptotene • Zygotene – synapsis of homologous chromosomes = bivalent Figure 3.8B: Zygotene • Pachytene – crossing-over between homologs Figure 3.8C: Early pachytene Parts A, B, and C courtesy of Marta Walters and Santa Barbara Botanic Gardens, Santa Barbara, California. Part D courtesy of Herbert Stern. Used with permission. Figure 3.8D: Late pachytene 27 Figure 3.9: Bivalent consisting of a pair of homologous chromosomes 28 Meiosis: Prophase I • Diplotene – chromosome repulsion, however, they remain held together by cross-connections resulting from crossing-over. Each cross-connection, called a chiasma, is formed by a breakage and rejoining between nonsister chromatids • Diakinesis – maximum chromosome contraction 29 Meiosis: Metaphase I • Metaphase I – the bivalents positioned with the centromeres of the two homologs on opposite sides of the metaphase plate • As each bivalent moves onto the metaphase plate, its centromeres are oriented at random with respect to the poles of the spindle • Genes on different chromosomes undergo independent assortment because nonhomologous chromosomes align at random in metaphase I 30 Figure 3.11: Independent assortment of genes on nonhomologous chromosomes 31 Meiosis: Anaphase I • Anaphase I – homologous chromosomes, each composed of two chromatids joined at an undivided centromere, separate from one another and move to opposite poles of the spindle • The physical separation of homologous chromosomes in anaphase I is the physical basis of Mendel’s principle of segregation 32 Meiosis: Telophase I • Telophase I – a haploid set of chromosomes consisting of one homolog from each bivalent is located near each pole of the spindle • The spindle breaks down, the chromosomes enter the second meiotic division after only a limited uncoiling • Chromosome replication never takes place between the two divisions 33 Meiosis II • The second meiotic division (meiosis II) is called the equational division because the chromosome number remains the same in each cell before and after the second division • In some species, the chromosomes pass directly from telophase I to prophase II without loss of condensation • After a short prophase II and the formation of second-division spindles, the centromeres of the chromosomes in each nucleus become aligned on the central plane of the spindle at metaphase II 34 Meiosis II • In anaphase II, the centromeres divide and the chromatids of each chromosome move to opposite poles of the spindle • Once the centromere has split at anaphase II, each chromatid is considered a separate chromosome • Telophase II is a transition to the interphase condition of the chromosomes in the four haploid nuclei, accompanied by division of the cytoplasm. 35 Meiosis • The chromatids of a chromosome are usually not genetically identical because of crossing-over associated with the formation of chiasmata during prophase of the first division 36 Figure 3.7: Chromosome behavior during meiosis 37 Chromosome Structure • Eukaryotic chromosomes are highly coiled stable complexes of DNA and protein called chromatin • Each eukaryotic chromosome contains a single DNA molecule of enormous length • Some of the proteins present in chromatin determine chromosome structure and the changes in structure during the cell cycle • Other chromatin proteins appear to have important roles in regulating chromosome functions 38 Chromatin Structure • The nucleosome is the basic structural unit of chromatin • Each nucleosome is composed of a core particle, ~55 base pairs of DNA called linker DNA that links adjacent core particles and one molecule of histone H1 that binds to the core particle and to the linker DNA • Histones are small proteins that are highly conserved among different organisms 39 Chromatin Structure • Each core particle consists of an octamere of pairs each of histone H2A, H2B, H3, and H4; a segment of DNA containing about 145 base pairs Figure 3.15a: Organization of nucleosomes 40 Figure 3.15b: Organization of nucleosomes 41 Chromatin Structure • In the nucleus of a nondividing cell, chromatin fibers form discrete chromosome territories • Chromosome territories are correlated with gene densities • Territories of chromosome domains that are relatively gene rich tend to be located toward the interior of the nucleus Figure courtesy of Tobias A. Knoch, Erasmus MC, Rotterdam, and Kirchhoff-Institute for Physics, Ruperto-Carola University, Heidelberg Figure 3.18: Chromosome territories formed by 30nm chromatin fibers within the nucleus of a nondividing cell 42 • Nucleosomes coil to form higher order DNA structure called the 30nm chromatin fiber Figure 3.19: Condensation of DNA and chromatin to form a metaphase chromosome 43 Chromatin Structure • The spaces between the chromatin domains form a network of channels large enough to allow passage of the molecular machinery for replication, transcription, and RNA processing • Replication takes place in small discrete regions that exhibit a reproducible temporal and spatial pattern, and transcription takes place in a few hundred discrete locations • The metaphase chromosome is a hierarchy of coiled coils 44 Chromatin Structure • Compact and heavily stained regions of chromatin are known as heterochromatin, which mainly consists of highly repeated noncoding DNA sequences—satellite DNA • The rest of the chromatin, which becomes visible only after chromosome condensation in mitosis or meiosis, is called euchromatin • The number of genes located in heterochromatin is small relative to the number in euchromatin 45 Figure 3.21a: Metaphase chromosomes of the ground squirrel Figure 3.21B: An interpretive drawing of metaphase chromosomes of the ground squirrel Part A courtesy of T.C. Hsu, Ph.D., and used with permission of Sen Pathak, Ph.D., Anderson Cancer Center, University of Texas. 46 Chromosome Structure • The centromere is essential for chromosome segregation • The centromere is a specific region of the eukaryotic chromosome. It serves as a central component of the kinetochore the complex of DNA and proteins to which the spindle fibers attach and move the chromosomes in both mitosis and meiosis 47 Figure 3.22: A yeast centromere Adapted from K. S. Bloom, M. Fitzgerald-Hayes, and J. Carbon, Cold Spring Harb. Symp. Quant. Biol. 47 (1982): 1175. 48 Chromosome Structure • The telomere is essential for the stability of the chromosome tips • Due to the nature of DNA replication, chromosomes require special mechanism to restore DNA in telomeres in each cycle of replication • The mechanism relies on an enzyme called telomerase 49 Figure 3.25: The function of telomerase 50 Figure 3.26: Telomere formation in Tetrahymena 51 Chromosomes and Heredity • Chromosome Theory of Heredity: Genes are located in chromosomes • Early evidence that genes are located on chromosomes was found by Thomas Hunt Morgan in 1910 • Morgan’s studied inheritance patterns in Drosophila melanogaster and found that in some cases reciprocal crosses yield different results 52 Morgan’s Fruit Fly Experiments • Morgan realized that it might happen if the alleles for some genes were present in the X chromosome • The X chromosome is transmitted in a different pattern by males and females, and the Y chromosome does not contain alleles homologous to genes on the X chromosome 53 Figure 3.29: A chromosomal interpretation of results obtained in F1 and F2 progenies in crosses of Drosophila 54 Nondisjunction • Experimental proof of the chromosome theory of heredity came from nondisjunction • Nondisjunction = chromosomes fail to separate (disjoin) and move to opposite poles of the division spindle, results in loss or gain of a chromosome • Calvin Bridges demonstrated that exceptional behavior of chromosomes is precisely paralleled by exceptional inheritance of their genes 55 Figure 3.32: The results of meiotic nondisjunction of the X chromosomes in a female Drosophila 56 X-Linked Inheritance • Special chromosomes determine sex in many organisms • X and Y chromosomes = sex chromosomes, which are nonidentical but share some genes • In most organisms, the Y chromosome carries few genes other than those related to male determination • X-linked genes are inherited according to sex • Hemophilia is a classic example of human X-linked inheritance 57 X-Linked Inheritance • In many organisms, the male is the heterogametic sex • Males produce two different types of gametes: one containing X and another Y chromosome • Females have two X chromosomes and produce only X-bearing gametes • In some organisms (birds, butterflies, and some reptiles), females are heterogametic 58 Data analysis • Genetic data analysis makes use of probability and statistics • Progeny of crosses are predicted by the binomial probability • If the probability of possibility A is p and the probability of the alternative possibility B is q, then the probability that, in n trials, A is realized s times and B is realized t times is n! psqt s!t! 59 Chi-Square Analysis • The test of goodness of fit = test analyzes whether observed data agree with theoretical expectation • A conventional measure of goodness of fit is a value called chi-square, c2 • c2 = ∑(observed – expected)2 / expected • A value of c2 = 0 means that the observed numbers fit the expected numbers perfectly 60 Chi-Square Analysis • Probability P that a worse fit (or one equally bad) would be obtained by chance, assuming that the genetic hypothesis is true • The critical values of P are conventionally chosen as 0.05 (the 5 percent level) and 0.01 (the 1 percent level) • Statistically significant refers to the magnitude of the difference between the observed and the expected numbers 61 Chi-Square Analysis • To determine the P value corresponding to a calculated c2 we need the number of degrees of freedom of the particular chi-square test • The number of degrees of freedom equals the number of classes of data minus 1 62 Figure 3.34: Graphs for interpreting goodness of fit to genetic predictions using the chi-square test 63