scholarly journals GENETIC ANALYSIS OF TWO ALLELIC TEMPERATURE-SENSITIVE MUTANTS OF DROSOPHILA MELANOGASTER BOTH OF WHICH ARE ZYGOTIC AND MATERNAL-EFFECT LETHALS

Genetics ◽  
1978 ◽  
Vol 89 (2) ◽  
pp. 341-353
Author(s):  
Allen Shearn ◽  
Grafton Hersperger ◽  
Evelyn Hersperger

ABSTRACT After fertilization, the development of a zygote depends upon both gene products synthesized by its maternal parent and gene products synthesized by the zygote itself. To analyze genetically the relative contributions of these two sources of gene products, several laboratories have been isolating two classes of mutants of Drosophila melanogaster: maternal-effect lethals and zygotic lethals. This report concerns the analysis of two temperature-sensitive mutants, OX736hs and PC025hs, which were isolated as alleles of a small-disc mutant, l(3)1902. These alleles are not only zygotic lethals, but also maternal-effect lethals. They have temperature-sensitive periods during larval life and during oogenesis. Mutant larvae exposed continuously to restrictive temperature have small discs. One- or two-day exposures to the restrictive temperature administered during the third larval instar lead to a homeotic transformation of the midlegs and hindlegs to the pattern characteristic of the forelegs. Mutant females exposed to the restrictive temperature during oogenesis produce eggs that can develop until gastrulation, but do not hatch. —The existence of these mutants, and one that was recently described by another group, implies that there may be a class of genes, heretofore unrecognized, whose products are synthesized during oogenesis, are essential for embryogenesis and are also synthesized during larval stages within imaginal disc cells.

Genetics ◽  
1993 ◽  
Vol 135 (2) ◽  
pp. 527-539 ◽  
Author(s):  
T R Parody ◽  
M A Muskavitch

Abstract Analysis of the development of Delta (Dl) temperature-sensitive mutants pulsed at restrictive temperature during larval and pupal stages reveals multiple phenocritical periods during which reduction of Dl function affects viability and development of adult structures. Dl function is required during the third larval instar for post-pupal viability and during the first day of pupal development for viability through eclosion. Dl function is required biphasically for the development of sensory bristles. Earlier pulses lead to bristle multiplication and later pulses lead to bristle loss. The exact intervals during which multiplication and loss are induced vary for different bristles. Dl function is also required for development of most, if not all, cell types in the retina. Different pulses result in reduction in eye size, scarring, and glossiness, as well as multiplication and loss of interommatidial bristles. We also define intervals during which Dl function is required for aspects of leg and wing development. Phenocritical periods for Dl function are temporally coincident with those previously reported for Notch (N), consistent with the hypothesis that the proteins encoded by Dl and N interact throughout development to assure correct specification of cell fates in a variety of imaginal tissues.


Genetics ◽  
1980 ◽  
Vol 96 (4) ◽  
pp. 939-965
Author(s):  
Jeffrey C Hall ◽  
Stamatis N Alahiotis ◽  
David A Strumpf ◽  
Kristin White

ABSTRACT Temperature-sensitive (ts) mutants of the Ace gene, which codes for acetylcholinesterase (AChE) in Drosophila melanogaster, were analyzed for defects in viability, behavior and function of the enzyme. The use of heat-sensitive and cold-sensitive mutations permited the function of AChE in the nervous system to be analyzed temporally. All ts mutations were lethal, or nearly so, when animals expressing them were subjected to restrictive temperatures during late embryonic and very early larval stages. Heat treatments to Ace-ts mid- and late larvae had little effect on the behavior of these animals or on the viability or behavior of the eventual adults. Heat-sensitive mutants exposed to nonpermissive temperatures as pupae, by contrast, had severe defects in phototaxis and locomotor activity as adults. AChE extracted from adult ts mutants that had developed at a permissive temperature were abnormally heat labile, and they had reduced substrate affinity when assayed at restrictive temperatures. However, enzyme activity did not decline during exposure of heat-sensitive adults to high temperatures even though such treatments caused decrements in phototaxis (29°) and, eventually, cessation of movement (31°). The cold-sensitive mutant also produced readily detectable levels of AChE when exposed to a restrictive temperature during the early developmental stage when this mutation causes almost complete lethality. We suggest that the relationship among the genetic, biochemical and neurobiological defects in these mutants may involve more than merely temperature-sensitive catalytic functions.


Genetics ◽  
1998 ◽  
Vol 148 (3) ◽  
pp. 989-1005 ◽  
Author(s):  
Keiko Umezu ◽  
Neal Sugawara ◽  
Clark Chen ◽  
James E Haber ◽  
Richard D Kolodner

Abstract Replication protein A (RPA) is a single-stranded DNA-binding protein identified as an essential factor for SV40 DNA replication in vitro. To understand the in vivo functions of RPA, we mutagenized the Saccharomyces cerevisiae RFA1 gene and identified 19 ultraviolet light (UV) irradiation- and methyl methane sulfonate (MMS)-sensitive mutants and 5 temperature-sensitive mutants. The UV- and MMS-sensitive mutants showed up to 104 to 105 times increased sensitivity to these agents. Some of the UV- and MMS-sensitive mutants were killed by an HO-induced double-strand break at MAT. Physical analysis of recombination in one UV- and MMS-sensitive rfa1 mutant demonstrated that it was defective for mating type switching and single-strand annealing recombination. Two temperature-sensitive mutants were characterized in detail, and at the restrictive temperature were found to have an arrest phenotype and DNA content indicative of incomplete DNA replication. DNA sequence analysis indicated that most of the mutations altered amino acids that were conserved between yeast, human, and Xenopus RPA1. Taken together, we conclude that RPA1 has multiple roles in vivo and functions in DNA replication, repair, and recombination, like the single-stranded DNA-binding proteins of bacteria and phages.


Temperature-sensitive mutants of polyoma virus have been examined to determine whether they are able to induce the synthesis of cellular DNA under conditions where viral gene products are defective. Two ‘early’ mutants, and one ‘late’ mutant of polyoma induce cellular DNA synthesis normally under conditions where virus growth is inhibited because viral gene products are defective.


Genetics ◽  
1979 ◽  
Vol 92 (4) ◽  
pp. 1079-1092
Author(s):  
Duane W Martindale ◽  
Ronald E Pearlman

ABSTRACT The parameters for the killing of Tetrahymena by 5-bromodeoxyuridine (BUdR) and near-ultraviolet light have been determined. Significant preferential killing by UV* of cells that have incorporated BUdR was obtained when the cells were irradiated in a nonnutrient buffer. UV alone was found to be toxic to cells irradiated in growth medium. Mutants defective in division at a restrictive temperature were isolated from mutagenized cultures that had been treated with BUdR and UV and from mutagenized cultures that had no such treatment. Results indicate that the number of temperature sensitive (ts) growth mutants can be increased five to six times using the BUdR/UV treatment. Data are presented that indicate differences in the frequency of occurrence of various types of ts mutants, with and without enrichment. A mutant that immediately stopped macromolecular synthesis and cell division upon being placed at the restrictive temperature was more resistant to BUdR/UV treatment than wild type by 1000-fold. Using the above techniques, BUdR-resistant mutants altered in the phosphorylation of thymidine have been isolated.


1976 ◽  
Vol 54 (12) ◽  
pp. 2089-2097 ◽  
Author(s):  
E. L. Peterson ◽  
J. D. Berger

One hundred and ninety-eight temperature-sensitive mutants of Paramecium tetraurelia were isolated after nitrosoguanidine mutagenesis. In some experiments, mutants were recovered with the aid of a bromouracil (BU) selection system. Fifty-six mutants showed cessation of cell division within one cell cycle at the restrictive temperature and were designated ts-0. Fourteen of the ts-0's showed a greater than 90% reduction in rnacronuclear deoxyribonucleic acid (DNA) synthesis at the restrictive temperature. Two ts-0. DNA-defective lines continued protein synthesis at greater than 50% the normal rate after arrest of DNA synthesis. Hence, these two mutants may be directly affected in the replication process itself. The two mutants are allelic and, in addition, a third 'leaky' allele was recovered. Comparison of experiments in which either BU selection or no selection was employed shows that a greater than 10-fold enrichment for ts mutants resulted from BU selection.


Development ◽  
1996 ◽  
Vol 122 (7) ◽  
pp. 2283-2294 ◽  
Author(s):  
P. Manfruelli ◽  
N. Arquier ◽  
W.P. Hanratty ◽  
M. Semeriva

Inactivation of the lethal(2)giant larvae (l(2)gl) gene results in malignant transformation of imaginal disc cells and neuroblasts of the larval brain in Drosophila. Subcellular localization of the l(2)gl gene product, P127, and its biochemical characterization have indicated that it participates in the formation of the cytoskeletal network. In this paper, genetic and phenotypic analyses of a temperature-sensitive mutation (l(2)glts3) that behaves as a hypomorphic allele at restrictive temperature are presented. In experimentally overaged larvae obtained by using mutants in the production of ecdysone, the l(2)glts3 mutation displays a tumorous potential. This temperature-sensitive allele of the l(2)gl gene has been used to describe the primary function of the gene before tumor progression. A reduced contribution of both maternal and zygotic activities in l(2)glts3 homozygous mutant embryos blocks embryogenesis at the end of germ-band retraction. The mutant embryos are consequently affected in dorsal closure and head involution and show a hypertrophy of the midgut. These phenotypes are accompanied by an arrest of the cell shape changes normally occurring in lateral epidermis and in epithelial midgut cells. l(2)gl activity is also necessary for larval fife and the critical period falls within the third instar larval stage. Finally, l(2)gl activity is required during oogenesis and mutations in the gene disorganize egg chambers and cause abnormalities in the shape of follicle cells, which are eventually internalized within the egg chamber. These results together with the tumoral phenotype of epithelial imaginal disc cells strongly suggest that the l(2)gl product is required in vivo in different types of epithelial cells to control their shape during development.


1984 ◽  
Vol 43 (3) ◽  
pp. 257-275 ◽  
Author(s):  
Donald A. R. Sinclair ◽  
Thomas A. Grigliatti ◽  
Thomas C. Kaufman

SUMMARYMinute(M) lesions exhibit a striking propensity for interacting with many different mutations. In the past, few attempts have been made to explain these diverse phenomena. This study describes a variety of temperature-sensitive (ts) interactions exhibited by the ts third chromosomeMinutemutationM(3)LS4Q-III(Q-III). Most of these interactions (i.e. those involvingvg, cp, Dl, DfdorLy) reflectQ-III-induced enhancement of the respective mutant phenotypes at the restrictive temperature. However,Q-IIIalso suppresses the extra-sex-comb phenotypes ofPcandMscat 29 °C and evokes lethal and bristle traits when combined withJ34eat the restrictive temperature. All of these interactions are characteristic of non-tsMinutelesions and thus they appear to be correlated with general physiological perturbations associated with theMsyndrome. In addition, our findings show that mutations that affect ribosome production and/or function, namelysu(f)ts67gandbbts−1, exhibit interactions comparable to those elicited byQ-III. Hence, in accordance with previous findings, we argue that most of theQ-IIIinteractions can be attributed to reduced translational capacity at the restrictive temperature. Finally, reciprocal temperature shift studies were used to delineate TSPs for interactions betweenQ-IIIandvg(mid to late second instar),cp(about mid-third instar),Dfd(early third instar) andDl(late second to mid third instar). We believe that these TSPs represent developmental intervals during which the respective gene products are utilized.


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