Jonathan Hodgkin

Evidence for evolutionary conservation of sex-determining genes

Most metazoans occur as two sexes. Surprisingly, molecular analyses have hitherto indicated that sex-determining mechanisms differ completely between phyla. Here we present evidence to the contrary. We have isolated the male sexual regulatory gene mab-3 (ref. 1) from the nematode Caenorhabditis elegans and found that it is related to the Drosophila melanogaster sexual regulatory gene doublesex (dsx). Both genes encode proteins with a DNA-binding motif that we have named the ‘DM domain’. Both genes control sex-specific neuroblast differentiation and yolk protein gene transcription; dsx controls other sexually dimorphic features as well. The form of DSX that is found in males can direct male-specific neuroblast differentiation in C. elegans. This structural and functional similarity between phyla suggests a common evolutionary origin of at least some aspects of sexual regulation. We have identified a human gene, DMT1, that encodes a protein with a DM domain and find that DMT1 is expressed only in testis. DMT1 maps to the distal short arm of chromosome 9, a location implicated in human XY sex reversal. Proteins with DM domains may therefore also regulate sexual development in mammals.

A Conserved Checkpoint Pathway Mediates DNA Damage–Induced Apoptosis and Cell Cycle Arrest in C. elegans

To maintain genomic stability following DNA damage, multicellular organisms activate checkpoints that induce cell cycle arrest or apoptosis. Here we show that genotoxic stress blocks cell proliferation and induces apoptosis of germ cells in the nematode C. elegans. Accumulation of recombination intermediates similarly leads to the demise of affected cells. Checkpoint-induced apoptosis is mediated by the core apoptotic machinery (CED-9/CED-4/CED-3) but is genetically distinct from somatic cell death and physiological germ cell death. Mutations in three genes--mrt-2, which encodes the C. elegans homolog of the S. pombe rad1 checkpoint gene, rad-5, and him-7-block both DNA damage-induced apoptosis and cell proliferation arrest. Our results implicate rad1 homologs in DNA damage-induced apoptosis in animals.

A uniform genetic nomenclature for the nematode Caenorhabditis elegans

SummaryA uniform system of genetic nomenclature for the nematode Caenorhabditis elegans is described. Convenient ways are specified to designate genes, mutations and strains, and to attempt to avoid name duplications.

mab-3, a gene required for sex-specific yolk protein expression and a male-specific lineage in C. elegans

The gene mab-3 appears to regulate a subset of sex-specific events in C. elegans male development. Mutations in mab-3 have no apparent effect on hermaphrodites, but cause synthesis of yolk proteins and a limited lineage alteration in males. We infer that mab-3 has at least two distinct male-specific functions. First, mab-3 activity prevents yolk protein production by males, without affecting stage or tissue specificity of expression. Second, mab-3 activity is required for expression of the male V ray cell lineage. Epistasis analysis is most consistent with a model in which mab-3 is controlled by tra-1, the last switch gene known to act in the somatic sex determination pathway. We discuss how genes such as mab-3 might generate sexual dimorphism.

More is Not Better: Brood Size and Population Growth in a Self-Fertilizing Nematode

The normal form of the nematode Caenorhabditis elegans is a self-fertilizing hermaphrodite, which produces from the same germ-line tissue first a limited number of sperm and then a larger number of oocytes. Self-progeny brood sizes are determined by the number of sperm, and most of the oocytes remain unfertilized. Therefore it might seem selectively advantageous to increase the number of sperm, and hence the size of the brood. A mutation that leads to a 50% increase in sperm production allows a comparison of population growth rates between the wild type (mean brood 327 progeny) and the mutant (mean brood 499 progeny). Wild-type populations grow faster, as measured by food consumption, indicating that increased brood size is not advantageous. The mutant appears to be at a disadvantage because the additional spermatogenesis leads to a delay in the onset of oogenesis, and hence to an increase in the minimum generation time. In support of the notion of an optimal brood size, it was found that different natural isolates of this species have self-fertilities similar to that of the standard laboratory strain, in the range 250—350 progeny per worm.

A sex-determining gene, fem-1, required for both male and hermaphrodite development in Caenorhabditis elegans.

Sex in the nematode Caenorhabditis elegans is normally determined by the X chromosome to autosome (X:A) ratio, with XX hermaphrodites and XO males. Previous work has shown that a set of at least four autosomal genes (her-1, tra-2, tra-3, and tra-1) is signaled by the X:A ratio and appears to act in a regulatory pathway to determine sex. Twenty-one new recessive alleles of the gene fem-1(IV) (formerly isx-1) have been isolated. Seven of these may be null alleles; one of these is an amber mutation. The other 14 alleles are temperature sensitive. The putative null mutations cause both XO and XX animals to develop as females when the mother as well as the zygote is fem-1(-). Therefore, fem-1(+) is required (a) for the development of the male body and (b) for spermatogenesis in males and hermaphrodites. In addition, fem-1 shows a maternal effect: wild-type fem-1 product partially rescues the development of fem-1(-) progeny. By analyzing double mutants it has been shown that fem-1(+) is part of the sex-determination pathway and has two distinct functions: (1) in the soma it prevents the action of tra-1, thereby allowing male development to occur, and (2) in the germline it is necessary for spermatogenesis in both sexes.

The product of fem-1, a nematode sex-determining gene, contains a motif found in cell cycle control proteins and receptors for cell-cell interactions

We report the cloning and sequencing of fem-1, a gene required for sex determination in both germline and somatic tissues in the nematode C. elegans. Clones carrying a 5.5 kb fragment are able to rescue the progeny of a fem-1 mutant when injected into its oocytes. The major fem-1 transcript in both sexes is 2.4 kb and comprises 11 exons. It encodes a soluble, intracellular protein of 656 amino acids that includes near its N-terminus six contiguous copies of a motif found in the products of the cdc10 gene of S. pombe, the SWI6 gene of S. cerevisiae, the Notch gene of Drosophila, and the lin-12 and glp-1 genes of C. elegans.

The Nematode Caenorhabditis elegans and Its Genome

Over the past two decades, the small soil nematode Caenorhabditis elegans has become established as a major model system for the study of a great variety of problems in biology and medicine. One of its most significant advantages is its simplicity, both in anatomy and in genomic organization. The entire haploid genetic content amounts to 100 million base pairs of DNA, about 1/30 the size of the human value. As a result, C. elegans has also provided a pilot system for the construction of physical maps of larger animal and plant genomes, and subsequently for the complete sequencing of those genomes. By mid-1995, approximately one-fifth of the complete DNA sequence of this animal had been determined. Caenorhabditis elegans provides a test bed not only for the development and application of mapping and sequencing technologies, but also for the interpretation and use of complete sequence information. This article reviews the progress so far toward a realizable goal-the total description of the genome of a simple animal.

X chromosome dosage and gene expression in Caenorhabditis elegans: Two unusual dumpy genes

SummaryThe phenotypes caused by mutations in two autosomal genes of the nematode Caenorhabditis elegans, dpy-21 V and dpy-26 IV, are markedly affected by X chromosome dosage, independent of sexual phenotype. At high X chromosome to autosome ratio, in 2A; 3X animals, these dumpy mutations are lethal; at intermediate ratio, in 2A; 2X animals, they cause dumpiness or lethality; at low ratio, in 2A; 1X animals they cause neither dumpiness nor lethality. One gene, dpy-26, exhibits a strong maternal effect. Interaction between these genes and two major sex-determining genes her-1 V and tra-1 III have been examined. The dumpy mutations partly suppress the masculinization of tra-1 2A;2X animals and also increase the fertility of most her-1 2A;1X hermaphrodites. It is suggested that these dumpy genes are involved in X chromosome dosage compensation, and in some aspects of sexual differentiation. The dpy-26 gene is compared with a similar Drosophila gene, daughterless.

Molecular identification of smg-4, required for mRNA surveillance in C.elegans

Premature termination codons trigger a process in eukaryotes known as nonsense mediated decay or mRNA surveillance, resulting in the rapid decay of the aberrant transcript. Studies in C. elegans have shown this system is mediated by seven smg genes and can prevent the accumulation of toxic, truncated peptides. Here we report the cloning of smg-4 by physical mapping and functional rescue assays. The minimal rescuing activity is found within a genomic operon, encoding a novel protein. The final exon of the gene is alternatively spliced for expression of two different isoforms. Although no known genes were found to exhibit significant homology to smg-4, a novel conserved domain has been identified by alignment with sequences defined by expressed sequence tags (ESTs) from a variety of organisms. Furthermore, we describe a homolog from C. briggsae, which will rescue C. elegans smg-4 mutants. The C. elegans gene has been fused to green fluorescent protein (GFP). This SMG-4:GFP fusion exhibits nuclear accumulation and diffuse cytoplasmic staining, and further localizes to what appear to be perinuclear and cytoplasmic punctate structures.