scholarly journals Inheritance of extrachromosomal ribosomal DNA during the asexual life cycle of Dictyostelium discoideum: examination by use of DNA polymorphisms.

1985 ◽  
Vol 5 (2) ◽  
pp. 273-280 ◽  
Author(s):  
D L Welker ◽  
K P Hirth ◽  
K L Williams
Keyword(s):  
Life Cycle ◽  
Dictyostelium Discoideum ◽  
Ribosomal Dna ◽  
Dna Polymorphisms ◽  
Haploid Cell ◽  
Wild Type ◽  
Parasexual Cycle ◽  
Restriction Fragments ◽  
Type Isolate ◽  
Ca 50

Wild-type isolates of Dictyostelium discoideum exhibited differences in the size of restriction fragments of the extrachromosomal 88-kilobase ribosomal DNA (rDNA) palindrome. Polymorphisms in rDNA also were found among strains derived solely from the NC4 wild-type isolate. These variations involved EcoRI fragments II, III, and V; they included loss of the EcoRI site separating fragments II and V and deletion and insertion of DNA. More than one rDNA form can coexist in the same diploid or haploid cell. However, one or another parental rDNA tended to predominate in diploids constructed, using the parasexual cycle, between haploid NC4-derived strains and haploid wild-type isolates. In some cases, most if not all of the rDNA of such diploids were of one form after ca. 50 generations of growth. Segregant haploids, derived from diploids that possessed predominantly a single rDNA allele, possessed the same allele as the diploid and did not recover the other form. This evidence implies that replication does not proceed from a single chromosomal or extrachromosomal copy of the rDNA during the asexual life cycle of D. discoideum.

Inheritance of extrachromosomal ribosomal DNA during the asexual life cycle of Dictyostelium discoideum: examination by use of DNA polymorphisms

1985 ◽  
Vol 5 (2) ◽  
pp. 273-280
Author(s):  
D L Welker ◽  
K P Hirth ◽  
K L Williams
Keyword(s):  
Life Cycle ◽  
Dictyostelium Discoideum ◽  
Ribosomal Dna ◽  
Dna Polymorphisms ◽  
Haploid Cell ◽  
Wild Type ◽  
Parasexual Cycle ◽  
Restriction Fragments ◽  
Type Isolate ◽  
Ca 50

Wild-type isolates of Dictyostelium discoideum exhibited differences in the size of restriction fragments of the extrachromosomal 88-kilobase ribosomal DNA (rDNA) palindrome. Polymorphisms in rDNA also were found among strains derived solely from the NC4 wild-type isolate. These variations involved EcoRI fragments II, III, and V; they included loss of the EcoRI site separating fragments II and V and deletion and insertion of DNA. More than one rDNA form can coexist in the same diploid or haploid cell. However, one or another parental rDNA tended to predominate in diploids constructed, using the parasexual cycle, between haploid NC4-derived strains and haploid wild-type isolates. In some cases, most if not all of the rDNA of such diploids were of one form after ca. 50 generations of growth. Segregant haploids, derived from diploids that possessed predominantly a single rDNA allele, possessed the same allele as the diploid and did not recover the other form. This evidence implies that replication does not proceed from a single chromosomal or extrachromosomal copy of the rDNA during the asexual life cycle of D. discoideum.

scholarly journals Mitochondrial processing peptidase activity is controlled by the processing of α-MPP during development in Dictyostelium discoideum

Microbiology ◽  
2010 ◽  
Vol 156 (4) ◽  
pp. 978-989 ◽  
Author(s):  
Koki Nagayama ◽  
Tetsuo Ohmachi
Keyword(s):  
Life Cycle ◽  
Dictyostelium Discoideum ◽  
Early Development ◽  
Normal Development ◽  
Peptidase Activity ◽  
Fruiting Bodies ◽  
Wild Type ◽  
Formation Stage ◽  
Mitochondrial Processing Peptidase ◽  
Processing Peptidase

We investigated the expression of the α subunit of the Dictyostelium mitochondrial processing peptidase (Ddα-MPP) during development. Ddα-MPP mRNA is expressed at the highest levels in vegetatively growing cells and during early development, and is markedly downregulated after 10 h of development. The Ddα-MPP protein is expressed as two forms, designated α-MPPH and α-MPPL, throughout the Dictyostelium life cycle. The larger form, α-MPPH, is cleaved to produce the functional α-MPPL form. We were not able to isolate mutants in which the α-mpp gene had been disrupted. Instead, an antisense transformant, αA2, expressing α-MPP at a lower level than the wild-type AX-3 was isolated to examine the function of the α-MPP protein. Development of the αA2 strain was normal until the slug formation stage, but the slug stage was prolonged to ∼24 h. In this prolonged slug stage, only α-MPPH was present, and α-MPPL protein and MPP activity were not detected. After 28 h, α-MPPL and MPP activity reappeared, and normal fruiting bodies were formed after a delay of approximately 8 h compared with normal development. These results indicate that MPP activity is controlled by the processing of α-MPPH to α-MPPL during development in Dictyostelium.

Cell sorting out during the differentiation of mixtures of metabolically distinct populations of Dictyostelium discoideum

Development ◽  
1973 ◽  
Vol 29 (3) ◽  
pp. 647-661
Author(s):  
C. K. Leach ◽  
J. M. Ashworth ◽  
D. R. Garrod
Keyword(s):  
Life Cycle ◽  
Dictyostelium Discoideum ◽  
Cell Sorting ◽  
Relative Number ◽  
Exponential Phase ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Axenic Medium ◽  
Spore Population ◽  
The Relationship

The behaviour, during the multicellular phase of the life-cycle, of amoebae of Dictyostelium discoideum grown in different media is described. Amoebal populations were marked by growth-temperature-sensitive genetic lesions which do not interfere with developmental phenomena. The fate of cell populations was determined by measuring the relative number of mutant and wild-type cells at various stages in the life-cycle. Cells sort out during development in such a way that they may be ordered in a sequence in which those given early in the following list preferentially appear in the spore population when mixed with those given later in the list: cells grown in axenic medium + 86 mm glucose and harvested when in the exponential phase of growth; cells grown in axenic medium and harvested when in the exponential phase of growth; cells grown on bacteria and harvested when in the exponential phase of growth; cells grown in axenic medium + 86 mM glucose and harvested when in the stationary phase of growth. Chemotactic aggregation and grex migration are not essential for sorting-out to occur but, in the normal life-cycle, the cells of a grex formed from amoebae grown in different media have sorted out anteroposteriorly. The relationship between this sorting out behaviour and the mechanism of pattern formation in fruiting-body morphogenesis is discussed. Differences in density of the amoebae cannot account for the sorting out predispositions we observe.

The use of a benomyl-resistant mutant to demonstrate latency of Colletotrichum gloeosporioides in avocado fruit

1993 ◽  
Vol 44 (4) ◽  
pp. 763 ◽  
Author(s):  
LM Coates ◽  
JAG Irwin ◽  
IF Muirhead
Keyword(s):  
Life Cycle ◽  
Ultraviolet Light ◽  
Growth Rates ◽  
Colletotrichum Gloeosporioides ◽  
Resistant Mutant ◽  
Field Conditions ◽  
Wild Type ◽  
Type Isolate ◽  
Avocado Fruit ◽  
First Time

A benomyl-resistant mutant of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc., generated by irradiating a wild-type isolate of the fungus with ultraviolet light, was used as a marker organism to demonstrate latency under field conditions. This mutant could be easily distinguished from wild-type isolates of C. gloeosporioides on the basis of growth rates on benomyl-amended media, and was as virulent in avocado fruit as wild-type isolates. Through the use of this mutant in field inoculations of avocado fruit, it was possible to demonstrate conclusively the existence, for the first time, of latency in the life cycle of C. gloeosporioides in this host. It was also shown that the fungus was able to remain latent for periods of at least 6 months.

scholarly journals Tandem repeats in extrachromosomal ribosomal DNA of Dictyostelium discoideum, resulting from chromosomal mutations.

Genetics ◽  
1992 ◽  
Vol 130 (4) ◽  
pp. 757-769
Author(s):  
R A Cole ◽  
K L Williams
Keyword(s):  
Linkage Group ◽  
Dictyostelium Discoideum ◽  
Ribosomal Dna ◽  
Tandem Repeats ◽  
Proximal Region ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Wild Type ◽  
Chromosomal Dna ◽  
Orthogonal Field

Abstract Extrachromosomal ribosomal DNA in the simple eukaryote Dictyostelium discoideum is readily separated from chromosomal DNA by orthogonal field electrophoresis (OFAGE), forming a prominent band in the 110-kb region of the gel. Here we show that mutations in at least two chromosomal genes give rise to a ladder of rDNA bands increasing in size up to about 300 kb. One of these mutations, the rrcA350 allele, which is recessive to wild type and maps to the centromere-proximal region of linkage group II, has an unstable phenotype; spontaneous revertants, which no longer exhibit the rDNA ladder, have been recovered. Another mutation rrc-351, provisionally mapped to linkage group IV, is dominant to wild type. The rDNA ladder is caused by concatamerization of a 34-kb fragment in the nontranscribed central spacer region of the 88-kb linear rDNA palindrome. Restriction enzyme analysis has revealed that each concatamer is generated by crossovers between two rDNA molecules.

scholarly journals Isolation of a gene required for programmed initiation of development by Aspergillus nidulans.

1992 ◽  
Vol 12 (9) ◽  
pp. 3827-3833 ◽  
Author(s):  
T H Adams ◽  
W A Hide ◽  
L N Yager ◽  
B N Lee
Keyword(s):  
Life Cycle ◽  
Aspergillus Nidulans ◽  
Optimal Growth ◽  
Growth Conditions ◽  
Nutrient Deprivation ◽  
Deletion Mutants ◽  
Wild Type ◽  
Microbial Development ◽  
Type Strains ◽  
Posttranscriptional Level

In contrast to many other cases in microbial development, Aspergillus nidulans conidiophore production initiates primarily as a programmed part of the life cycle rather than as a response to nutrient deprivation. Mutations in the acoD locus result in "fluffy" colonies that appear to grow faster than the wild type and proliferate as undifferentiated masses of vegetative cells. We show that unlike wild-type strains, acoD deletion mutants are unable to make conidiophores under optimal growth conditions but can be induced to conidiate when growth is nutritionally limited. The requirement for acoD in conidiophore development occurs prior to activation of brlA, a primary regulator of development. The acoD transcript is present both in vegetative hyphae prior to developmental induction and in developing cultures. However, the effects of acoD mutations are detectable only after developmental induction. We propose that acoD activity is primarily controlled at the posttranscriptional level and that it is required to direct developmentally specific changes that bring about growth inhibition and activation of brlA expression to result in conidiophore development.

scholarly journals DIFFERENTIATION OF DROSOPHILA CELLS LACKING RIBOSOMAL DNA, IN VITRO

Genetics ◽  
1973 ◽  
Vol 73 (3) ◽  
pp. 429-434
Author(s):  
J James Donady ◽  
R L Seecof ◽  
M A Fox
Keyword(s):  
Drosophila Melanogaster ◽  
Ribosomal Rna ◽  
Ribosomal Dna ◽  
Rna Synthesis ◽  
Wild Type ◽  
Drosophila Cells

ABSTRACT Drosophila melanogaster embryos that lacked ribosomal DNA were obtained from appropriate crosses. Cells were taken from such embryos before overt differentiation took place and were cultured in vitro. These cells differentiated into neurons and myocytes with the same success as did wild-type controls. Therefore, ribosomal RNA synthesis is not necessary for the differentiation of neurons and myocytes in vitro.

scholarly journals Borrelia burgdorferi Lacking BBK32, a Fibronectin-Binding Protein, Retains Full Pathogenicity

2006 ◽  
Vol 74 (6) ◽  
pp. 3305-3313 ◽  
Author(s):  
Xin Li ◽  
Xianzhong Liu ◽  
Deborah S. Beck ◽  
Fred S. Kantor ◽  
Erol Fikrig
Keyword(s):  
Life Cycle ◽  
Borrelia Burgdorferi ◽  
Deletion Mutant ◽  
Binding Protein ◽  
Kanamycin Resistance ◽  
Binding Activity ◽  
Mammalian Host ◽  
Wild Type ◽  
Fibronectin Binding Protein ◽  
Fibronectin Binding

ABSTRACT BBK32, a fibronectin-binding protein of Borrelia burgdorferi, is one of many surface lipoproteins that are differentially expressed by the Lyme disease spirochete at various stages of its life cycle. The level of BBK32 expression in B. burgdorferi is highest during infection of the mammalian host and lowest in flat ticks. This temporal expression profile, along with its fibronectin-binding activity, strongly suggests that BBK32 may play an important role in Lyme pathogenesis in the host. To test this hypothesis, we constructed an isogenic BBK32 deletion mutant from wild-type B. burgdorferi B31 by replacing the BBK32 gene with a kanamycin resistance cassette through homologous recombination. We examined both the wild-type strain and the BBK32 deletion mutant extensively in the experimental mouse-tick model of the Borrelia life cycle. Our data indicated that B. burgdorferi lacking BBK32 retained full pathogenicity in mice, regardless of whether mice were infected artificially by syringe inoculation or naturally by tick bite. The loss of BBK32 expression in the mutant had no adverse effect on spirochete acquisition (mouse-to-tick) and transmission (tick-to-mouse) processes. These results suggest that additional B. burgdorferi proteins can complement the function of BBK32, fibronectin binding or otherwise, during the natural spirochete life cycle.

Investigation of a potential role for aldose reductase AlrA in tetrahydropteridine synthesis in Dictyostelium discoideum Ax2

Pteridines ◽  
2017 ◽  
Vol 28 (2) ◽  
pp. 97-103
Author(s):  
Hye-Lim Kim ◽  
Hyun-Chul Ryu ◽  
Young Shik Park
Keyword(s):  
Dictyostelium Discoideum ◽  
Aldose Reductase ◽  
Potential Role ◽  
Amino Acid Residues ◽  
Wild Type ◽  
High Sequence Identity ◽  
Physiological Conditions ◽  
Antioxidant Function ◽  
Cellular Extract ◽  
Sr Gene

AbstractDictyostelium discoideum Ax2 is well-known for the synthesis of d-threo-tetrahydrobiopterin (DH4) with a smaller amount of l-erythro-tetrahydrobiopterin (BH4). DH4 synthesis from 6-pyruvoyltetrahydropterin (PPH4) is catalyzed by aldose reductase (AR)-like protein and sepiapterin reductase (SR) via an intermediate 1′-oxo-2′-d-hydroxypropyl tetrahydropterin, which is non-enzymatically oxidized to d-sepiapterin in the absence of SR. However, l-sepiapterin was a dominant product in the reaction of a cellular extract of spr− disrupted in the SR gene. In order to investigate its potential role in tetrahydropteridine synthesis, the enzyme catalyzing l-sepiapterin synthesis from PPH4 was purified from spr−. Via mass spectrometry, the protein was identified to be encoded by alrA. AlrA consists of 297 amino acid residues sharing a high sequence identity with human AR. However, in the co-incubation assay, DH4 synthesis was not detected and, furthermore, the recombinant AlrA was observed to suppress BH4 synthesis by SR, which was known to prefer 1′-oxo-2′-d-hydroxypropyl tetrahydropterin to PPH4. Although intracellular DH4 level in alrA− was decreased to 60% of the wild type, it is presumed to result from the antioxidant function of DH4. Therefore, despite the structural and catalytic identities with human AR, AlrA seems to be involved in neither BH4, nor DH4 synthesis under normal physiological conditions.

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