ABSTRACT
Faithful chromosome inheritance is a fundamental biological activity and errors contribute to birth defects and cancer progression. We have performed a P-element screen in Drosophila melanogaster with the aim of identifying novel candidate genes involved in inheritance. We used a "sensitized" minichromosome substrate (J21A) to screen approximately 3,000 new P-element lines for dominant effects on chromosome inheritance and recovered 78 Sensitized chromosome inheritance modifiers (Scim). Of these, 69 decreased minichromosome inheritance while 9 increased minichromosome inheritance. Fourteen mutations are lethal or semilethal when homozygous and all exhibit dramatic mitotic defects. Inverse PCR combined with genomic analyses identified P insertions within or close to genes with previously described inheritance functions, including wings apart-like (wapl), centrosomin (cnn), and pavarotti (pav). Further, lethal insertions in replication factor complex 4 (rfc4) and GTPase-activating protein 1 (Gap1) exhibit specific mitotic chromosome defects, discovering previously unknown roles for these proteins in chromosome inheritance. The majority of the lines represent mutations in previously uncharacterized loci, many of which have human homologs, and we anticipate that this collection will provide a rich source of mutations in new genes required for chromosome inheritance in metazoans.
Subject(s)
Cytoskeletal Proteins , Drosophila Proteins , Drosophila melanogaster/genetics , Genes, Insect , Microfilament Proteins , RNA-Binding Proteins , Transcription Factors , Animals , Checkpoint Kinase 1 , DNA Transposable Elements , Eukaryotic Initiation Factor-4E , Eye Proteins/genetics , Female , Glycoproteins/genetics , Histones/genetics , Homeodomain Proteins/genetics , Insect Proteins/genetics , Male , Membrane Glycoproteins/genetics , Microtubule-Associated Proteins/genetics , Peptide Initiation Factors/genetics , Protein Kinases/genetics , Trans-Activators/genetics , rab5 GTP-Binding Proteins/genetics , ras GTPase-Activating Proteins/geneticsABSTRACT
Pulsed-field gel analysis of Magnetospirillum magnetotacticum, strain MS-1, indicates that the genome is a single, circular structure of about 4.3 mb. A few genes, identified by sequence similarity, have been localized and arranged in a map with dnaA, indicating the presumed origin of replication. There are at least two rRNA operons. In addition, rRNA genes are found on a 40 kb, possibly extrachromosomal, structure. The genes thought to be involved in magnetite synthesis, bfr and magA, are located in the same 17% of the genome. A one base pair-overlap seen in the bfr genes of MS-1 is found also in the closely related magnetic strain AMB-1, but not in the non-magnetic relative A. itersonii.
