Annual Review of Genetics - Volume 31, 1997

Production of red blood cells (erythropoiesis) in the vertebrate embryo is critical to its survival and subsequent development. As red cells are the first blood cells to appear in embryogenesis, their origin reflects commitment of mesoderm to an hematopoietic fate and provides an avenue by which to examine the development of the hematopoietic system, including the hematopoietic stem cell (HSC). We discuss the genetics of erythropoiesis as studied in two systems: the mouse and zebrafish (Danio rerio). In the mouse, targeted disruption has established several genes as essential at different stages of hematopoiesis or erythroid precursor cell maturation. In the zebrafish, numerous mutants displaying a wide range of phenotypes have been isolated, although the affected genes are unknown. In comparing mouse knockout and zebrafish mutant phenotypes, we propose a pathway for erythropoiesis that emphasizes the apparent similarity of the mutants and the complementary nature of investigation in the two species. We speculate that further genetic studies in mouse and zebrafish will identify the majority of essential genes and define a regulatory network for hematopoiesis in vertebrates.

During this decade, there have been major advancements in the understanding of genetic loci involved in synthesis of the family of Mg-tetrapyrroles known as chlorophylls and bacteriochlorophylls. Molecular genetic analysis of Mg-tetrapyrrole biosynthesis was initiated by the performance of detailed sequence and mutational analysis of the photosynthesis gene cluster from Rhodobacter capsulatus. These studies provided the first detailed understanding of genes involved in bacteriochlorophyll a biosynthesis. In the short time since these studies were initiated, most of the chlorophyll biosynthesis genes have been identified by virtue of their ability to complement bacteriochlorophyll a biosynthesis mutants as well as by sequence homology comparisons. This review is centered on a discussion of our current understanding of bacterial, algal, and plant genes that code for enzymes in the Mg-branch of the tetrapyrrole biosynthetic pathway that are responsible for synthesis of chlorophylls and bacteriochlorophylls.

Gene amplification is a common feature of the genome of prokaryotic organisms. In this review, we analyze different instances of gene amplification in a variety of prokaryotes, including their mechanisms of generation and biological role. Growing evidence supports the concept that gene amplification be considered not as a mutation but rather as a dynamic genomic state related to the adaptation of bacterial populations to changing environmental conditions or biological interactions. In this context, the potentially amplifiable DNA regions impose a defined dynamic structure on the genome. If such structure has indeed been selected during evolution, it is a particularly challenging hypothesis.

Listeria monocytogenes is a pathogenic bacterium that induces its own uptake into mammalian cells, and spreads from one cell to another by an actin-based motility process. Entry into host cells involves the bacterial surface proteins InlA (internalin) and InlB. The receptor for InlA is the cell adhesion molecule E-cadherin. InlB-mediated entry requires activation of the host protein phosphoinositide (PI) 3-kinase, probably in response to engagement of a receptor. Actin-based movement of L. monocytogenes is mediated by the bacterial surface protein ActA. The N-terminal region of this protein is necessary and sufficient for polymerization of host cell actin. Other host proteins involved in bacterial motility include profilin, Vasodilator-Stimulated Phosphoprotein (VASP), the Arp2/Arp3 complex, and cofilin. Studies of entry and intracellular movement of L. monocytogenes could lead to a better understanding of receptor-ligand signaling and dynamics of actin polymerization in mammalian cells.

Prions are unprecedented infectious pathogens that cause a group of invariably fatal, neurodegenerative diseases by an entirely novel mechanism. Prion diseases may present as genetic, infectious, or sporadic disorders, all of which involve modification of the prion protein (PrP). The human prion disease Creutzfeldt-Jakob disease (CJD) generally presents as a progressive dementia, whereas scrapie of sheep and bovine spongiform encephalopathy (BSE) are manifest as ataxic illnesses. Prions are devoid of nucleic acid and seem to be composed exclusively of a modified isoform of PrP designated PrP^Sc. The normal, cellular PrP designated PrP^C is converted into PrP^Sc through a process whereby some of its α-helical structure is converted into β-sheet. The species of a particular prion is encoded by the sequence of the chromosomal PrP gene of the mammals in which it last replicated. In contrast to pathogens with a nucleic acid genome, prions encipher strain-specific properties in the tertiary structure of PrP^Sc. Transgenetic studies argue that PrP^Sc acts as a template upon which PrP^C is refolded into a nascent PrP^Sc molecule through a process facilitated by another protein.

Many viruses utilize the cellular transcription apparatus to express their genomes, and they encode transcriptional regulatory proteins that modulate the process. Here we review the current understanding of three viral regulatory proteins. The adenovirus E1A protein acts within the nucleus to regulate transcription through its ability to bind to other proteins. The herpes simplex type 1 virus VP16 protein acts within the nucleus to control transcription by binding to DNA in conjunction with cellular proteins. The human T-cell leukemia virus Tax protein influences transcription through interactions with cellular proteins in the nucleus as well as the cytoplasm.

The RuvA, RuvB, and RuvC proteins in Escherichia coli play important roles in the late stages of homologous genetic recombination and the recombinational repair of damaged DNA. Two proteins, RuvA and RuvB, form a complex that promotes ATP-dependent branch migration of Holliday junctions, a process that is important for the formation of heteroduplex DNA. Individual roles for each protein have been defined, with RuvA acting as a specificity factor that targets RuvB, the branch migration motor, to the junction. Structural studies indicate that two RuvA tetramers sandwich the junction and hold it in an unfolded square-planar configuration. Hexameric rings of RuvB face each other across the junction and promote a novel dual helicase action that “pumps” DNA through the RuvAB complex, using the free energy provided by ATP hydrolysis. The third protein, RuvC endonuclease, resolves the Holliday junction by introducing nicks into two DNA strands. Genetic and biochemical studies indicate that branch migration and resolution are coupled by direct interactions between the three proteins, possibly by the formation of a RuvABC complex.

Mating type genes regulate sexual compatibility and sexual reproduction in fungi. This review focuses on recent molecular analyses of well-characterized mating systems from representative ascomycete (Neurospora crassa, Podospora anserina) and basidiomycete (Ustilago maydis, Coprinus cinereus, Schizophyllum commune) fungi. These mating systems include many conserved components, such as gene regulatory polypeptides and pheromone/receptor signal transduction cascades, as well as conserved processes, like self-nonself recognition and controlled nuclear migration. The components' structures and their genetic arrangements in the mating system vary greatly in different fungi. Although similar components and processes are also found in ascomycete yeasts (Saccharomyces cerevisiae and Schizosaccharomyces pombe), the filamentous systems exhibit properties not encountered in yeast. Mating type genes act within, and control the development of, spatially differentiated fruiting bodies. The complex mating systems of basidiomycetes, unlike ascomycete systems, involve novel one-to-many specificity in both pheromone-receptor and homeodomain protein interactions.

Eukaryotic cells evolved when their genetic information was packed into the cell nucleus. DNA replication and RNA biogenesis occur inside the nucleus while protein synthesis takes place in the cytoplasm. Bi-directional trafficking between these two compartments is mediated by a single supramolecular assembly, the nuclear pore complex. Nucleocytoplasmic transport is signal mediated, energy dependent, and requires, besides nuclear pore proteins (nucleoporins), a number of soluble transport factors. We review here our current knowledge on the role of nucleoporins, and on the mechanism of nucleocytoplasmic transport, with emphasis on the yeast Saccharomyces cerevisiae.

A central unanswered question in phototransduction is how photosensitive molecules, visual pigments, regulate their absorption spectra. In nature, there exist various types of visual pigments that are adapted to diverse photic environments. To elucidate the molecular mechanisms involved in the adaptive selection of these pigments, we have to identify amino acid changes of pigments that are potentially important in changing the wavelength of maximal absorption (λmax) and then determine the effects of these mutations on the shift in λmax. The desired mutants can be constructed using site-directed mutagenesis, expressed in tissue culture cells, and the functional effect of virtually any such mutant can be rigorously determined. The availability of these cell/molecular methods makes vision an ideal model system in studying adaptive mechanisms at the molecular level. The identification of potentially important amino acid changes using evolutionary biological means is an indispensable step in elucidating the molecular mechanisms that underlie the spectral tuning of visual pigments.

The mariner/Tc1 superfamily of transposable elements is one of the most diverse and widespread Class II transposable elements. Within the larger assemblage, the mariner-like elements (MLEs) and the Tc1-like elements (TLEs) are distinct families differing characteristically in the composition of the “D,D(35)E” cation-binding domain. Based on levels of sequence similarity, the elements in each family can be subdivided further into several smaller subfamilies. MLEs and TLEs both have an extraordinarily wide host range. They are abundant in insect genomes and other invertebrates and are found even in some vertebrate species including, in the case of mariner, humans, in which one element on chromosome 17p has been implicated as a hotspot of recombination. In spite of the extraordinary evolutionary success of the elements, virtually nothing is known about their mode of regulation within genomes. There is abundant evidence that the elements are disseminated to naive host genomes by horizontal transmission, and there is a substantial base of evidence for inference about the subsequent population dynamics. Studies of engineered mariner elements and induced mutations in the transposase have identified two mechanisms that may be operative in mariner regulation. One mechanism is overproduction inhibition, in which excessive wild-type transposase reduces the rate of excision of a target element. A second mechanism is dominant-negative complementation, in which certain mutant transposase proteins antagonize the activity of the wild-type transposase. The latter process may help explain why the vast majority of MLEs in nature undergo “vertical inactivation” by multiple mutations and, eventually, stochastic loss. There is also evidence that mariner/Tc1 elements can be mobilized in hybrid dysgenesis; in particular, certain dysgenic crosses in Drosophila virilis result in mobilization of a TLE designated Paris as well as the mobilization of other unrelated transposable elements.

Volvox carteri possesses only two cell types: mortal somatic cells and potentially immortal asexual reproductive cells called gonidia. Mutational analysis indicates that three categories of genes play central roles in programming this germ-soma division of labor: First the gls genes function during embryogenesis to cause asymmetric divisions that produce large and small cells. Then the lag genes act in the large cells (gonidial initials) to repress functions required for somatic development while the regA locus acts in the small cells (somatic initials) to repress functions required for reproductive development. Transposon tagging and DNA transformation have recently been used to recover and characterize the glsA and regA genes, and the sequences of these genes lead to testable hypotheses about how they play their roles in germ-soma differentiation.

Transposable elements propagate by inserting into new locations in the genomes of the hosts they inhabit. Their transposition might thus negatively affect the fitness of the host, suggesting the requirement for a tight control in the regulation of transposable element mobilization. The nature of this control depends on the structure of the transposable element. DNA elements encode a transposase that is necessary, and in most cases sufficient, for mobilization. In general, regulation of these elements depends on intrinsic factors with little direct input from the host. Retrotransposons require an RNA intermediate for transposition, and their frequency of mobilization is controlled at multiple steps by the host genome by regulating both their expression levels and their insertional specificity. As a result, a symbiotic relationship has developed between transposable elements and their host. Examples are now emerging showing that transposons can contribute significantly to the well being of the organisms they populate.

In a wide variety of organisms, gametes develop within clusters of interconnected germline cells called cysts. Four major principles guide the construction of most cysts: synchronous division, a maximally branched pattern of interconnection between cells, specific changes in cyst geometry, and cyst polarization. The fusome is a germline-specific organelle that is associated with cyst formation in many insects and is likely to play an essential role in these processes. This review examines the cellular and molecular processes that underlie fusome formation and cyst initiation, construction, and polarization in Drosophila melanogaster. The studies described here highlight the importance of cyst formation to the subsequent development of functional gametes.

This article highlights recent advances in the molecular structure and function of proteins that are activated or created by chromosomal abnormalities and discusses their possible role in tumor development. The molecular characterization of these proteins has revealed that tumor-specific fusion proteins are the consequence of most chromosome translocations associated with leukemias and solid tumors. An emerging common theme is that creation of these proteins disrupts the normal development of tumor-specific target cells by blocking apoptosis. These insights identify these chromosomal translocation-associated genes as potential targets for improved cancer therapies.

Germline stem cells (GSCs) are the self-renewing population of germ cells that serve as the source for gametogenesis. GSCs exist in diverse forms, from those that undergo strict self-renewing asymmetric divisions in Drosophila to those that maintain their population by balancing between mitosis and differentiation in Caenorhabditis elegans. Most vertebrate spermatogonial GSCs appear to adopt an intermediate strategy. In most animals, GSCs are established during preadult gonadogenesis following the proliferation and migration of embryonic primordial germ cells. GSCs produce numerous gametes throughout the sexually active period of adult life. The establishment and self-renewing division of GSCs are controlled by extracellular signals such as hormones from the hypothalamic-pituitary axis and local interactions between GSCs and their neighboring cells. These extracellular signals may then influence differential gene expression, cell cycle machinery, and cytoskeletal organization of GSCs for their formation and/or divisional asymmetry. In addition, the GSC mechanism is related to that for germline and sex determination. Current knowledge has provided a solid framework for further study of GSCs and stem cells in general.

A handful of autosomal genes in the mammalian genome are inherited in a silent state from one of the two parents, and in a fully active form from the other, thereby rendering the organism functionally hemizygous for imprinted genes. To date 19 imprinted genes have been identified; 5 are expressed from the maternal chromosome while the rest are expressed from the paternal chromosome. Allele-specific methylation of CpG residues, established in one of the germlines and maintained throughout embryogenesis, has been clearly implicated in the maintenance of imprinting in somatic cells. Although the function of imprinting remains a subject of some debate, the process is thought to have an important role in regulating the rate of fetal growth.

The general goal of genetic studies of learning and memory is to develop and test theories that explain the animal's behavior in neuroanatomical, neurophysiological, cellular, and molecular terms. In this review we describe the role that gene targeting and other transgenic techniques have had in the study of mammalian learning and memory. We focus especially on the hippocampus, a brain structure that is thought to be central to the processing and temporary storage of complex information. We also discuss the main issues that confront this young field, as well as our vision for its future.

Arabidopsis thaliana (Arabidopsis) is proving to be an ideal model system for studies of host defense responses to pathogen attack. The Arabidopsis genetic system is significantly more tractable than those of other plant species, and Arabidopsis exhibits all of the major kinds of defense responses described in other plants. A large number of virulent and avirulent bacterial, fungal, and viral pathogens of Arabidopsis have been collected. In the last few years, a large number of mutations have been identified in Arabidopsis that cause a wide variety of specific defense-related phenotypes. Analysis of these mutant phenotypes is beginning to give glimpses into the complex signal transduction pathways leading to the induction of the defense responses involved in protecting plants from pathogen infection.