scholarly journals Functional Analysis of Deletion Derivatives of the Maize Transposon MuDR Delineates Roles for the MURA and MURB Proteins

Genetics ◽  
1999 ◽  
Vol 151 (1) ◽  
pp. 331-341 ◽  
Author(s):  
Damon Lisch ◽  
Lisa Girard ◽  
Maureen Donlin ◽  
Michael Freeling
Keyword(s):  
Functional Analysis ◽  
Transposable Elements ◽  
Antisense Transcript ◽  
Northern Analysis ◽  
Deletion Derivative ◽  
Mutator Activity ◽  
Sequences Analysis ◽  
Derivatives Of

Abstract The regulatory transposon of the Mutator system of transposable elements in maize is MuDR. MuDR elements produce two transcripts, from genes mudrA and mudrB, encoding proteins MURA and MURB, respectively. Like many other transposons, MuDR elements often undergo deletions, usually of internal sequences. Analysis of a deletion that is restricted to the region encoding MURB demonstrates that this gene is not required to cause excisions of a reporter element, although it may be required for transposition or suppression of suppressible alleles. Conversely, a derivative that lacks the region encoding MURA but that produces MURB is nonfunctional for all aspects of Mutator activity. Northern analysis of these derivatives reveals that each of the two transcripts can be independently transcribed, and analysis using an antibody specific for MURB reveals that mudrB transcript can also be successfully translated and its product appropriately localized in the absence of mudrA. A third deletion derivative provides evidence for a source of previously reported antisense transcript.

scholarly journals Hfr FORMATION DIRECTED BY Tn10

Genetics ◽  
1979 ◽  
Vol 91 (4) ◽  
pp. 639-655
Author(s):  
Forrest G Chumley ◽  
Rolf Menzel ◽  
John R Roth
Keyword(s):  
Drug Resistance ◽  
Homologous Recombination ◽  
Genetic Mapping ◽  
Transposable Elements ◽  
Salmonella Typhimurium ◽  
F Plasmid ◽  
Chromosome Transfer ◽  
Derivatives Of ◽  
Bacterial Systems

ABSTRACT The transposable drug-resistance element, TnlO, can serve as a region of homology to direct the insertion of an F′tsll4 lac plasmid into the chromosome of Salmonella typhimurium. Derivatives of F′tsl14 lac were constructed that carry TnlO insertions; these plasmids were transferred to strains having a TnlO insertion in the chromosome. Under these circumstances, Hfr formation requires homologous recombination between plasmid-borne and Chromosomal TnlO elements. The process is dependent on recA function and on the presence of both TnlO elements. All Hfr's isolated from a given merodiploid show the same direction of transfer. Depending on the orientation of TnlO in the F′ plasmid, Hfr's transferring in either direction can be obtained from any chromosomal TnlO insertion. Since TnlO insertions can be generated in any region of the chromosome, this method permits the isolation of Hfr's with either direction of transfer having their origin at almost any predetermined site. The Hfr's constructed by this method are sufficiently stable for standard genetic mapping crosses, and they have also been used to generate new F′ plasmids. Implicit in the results above is the possibility of determining the orientation of any chromosomal TnlO insertion by constructing an Hfr using a standard F′ TnlO plasmid and determining the direction of chromosome transfer. The general approaches described here are applicable to other transposable elements and other bacterial systems.

scholarly journals Cloning of the Mutator transposable element MuA2, a putative regulator of somatic mutability of the a1-Mum2 allele in maize.

Genetics ◽  
1991 ◽  
Vol 129 (3) ◽  
pp. 845-854 ◽  
Author(s):  
M M Qin ◽  
D S Robertson ◽  
A H Ellingboe
Keyword(s):  
Transposable Elements ◽  
Transposable Element ◽  
Copy Number ◽  
Inverted Repeats ◽  
Terminal Inverted Repeats ◽  
Somatic Instability ◽  
Mutator Activity ◽  
Element System ◽  
Lines Of Evidence

Abstract The identification of the autonomous or transposase-encoding element of the Mutator (Mu) transposable element system of maize is necessary to the characterization of the system. We reported previously that a transcript homologous to the internal region of the MuA element is associated with activity of the Mutator system. We describe here the cloning of another Mu element, designated MuA2, that cosegregates with Mutator activity as assayed by somatic instability of the a1-Mum2 allele. The MuA2 element has features typical of the transposable elements of the Mutator family, including the 210-bp terminal inverted repeats. Several lines of evidence suggest that MuA2 is an autonomous or transposase-encoding element of the Mu family: (1) MuA2 cosegregates with a genetically defined element that regulates somatic mutability of the a1-Mum2 allele; (2) MuA2 is hypomethylated while most other MuA2-hybridizing sequences in the genome are extensively methylated; (3) the increase of the copy number of MuA2 is concomitant with the increase of regulator elements; (4) MuA2-like elements are found in Mutator lines but not in non-Mutator inbreds. We propose that autonomous or transposase-encoding elements of the Mu family may be structurally conserved and MuA2-like.

scholarly journals Gentisate and 3-oxoadipate pathways in the yeast Candida parapsilosis: identification and functional analysis of the genes coding for 3-hydroxybenzoate 6-hydroxylase and 4-hydroxybenzoate 1-hydroxylase

Microbiology ◽  
2011 ◽  
Vol 157 (7) ◽  
pp. 2152-2163 ◽  
Author(s):  
Zuzana Holesova ◽  
Michaela Jakubkova ◽  
Ivana Zavadiakova ◽  
Igor Zeman ◽  
Lubomir Tomaska ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Candida Parapsilosis ◽  
Intracellular Localization ◽  
Cultivation Medium ◽  
Pathogenic Yeast ◽  
Gentisate Pathway ◽  
Derivatives Of

The pathogenic yeast Candida parapsilosis degrades various hydroxy derivatives of benzenes and benzoates by the gentisate and 3-oxoadipate pathways. We identified the genes MNX1, MNX2, MNX3, GDX1, HDX1 and FPH1 that code for enzymes involved in these pathways in the complete genome sequence of C. parapsilosis. Next, we demonstrated that MNX1, MNX2, MNX3 and GDX1 are inducible and transcriptionally controlled by hydroxyaromatic substrates present in cultivation media. Our results indicate that MNX1 and MNX2 code for flavoprotein monooxygenases catalysing the first steps in the 3-oxoadipate and gentisate pathways, respectively (i.e. 4-hydroxybenzoate 1-hydroxylase and 3-hydroxybenzoate 6-hydroxylase). Moreover, we found that the two pathways differ by their intracellular localization. The enzymes of the 3-oxoadipate pathway, Mnx1p and Mnx3p, localize predominantly in the cytosol. In contrast, intracellular localization of the components of the gentisate pathway, Mnx2p and Gdx1p, depends on the substrate in the cultivation medium. In cells growing on glucose these proteins localize in the cytosol, whereas in media containing hydroxyaromatic compounds they associate with mitochondria. Finally, we showed that the overexpression of MNX1 or MNX2 increases the tolerance of C. parapsilosis cells to the antifungal drug terbinafine.

scholarly journals Transcriptional analysis of Ty1 deletion and inversion derivatives at CYC7

1986 ◽  
Vol 6 (10) ◽  
pp. 3299-3311
Author(s):  
M Company ◽  
B Errede
Keyword(s):  
Cytochrome C ◽  
Transcriptional Activation ◽  
Fold Increase ◽  
Transcriptional Analysis ◽  
Insertion Mutation ◽  
Adjacent Gene ◽  
Coding Region ◽  
Deletion Derivative ◽  
And Inversion ◽  
Derivatives Of

One class of Ty insertion mutation in Saccharomyces cerevisiae activates expression of adjacent structural genes. The CYC7-H2 mutation, in which a Ty1 element is inserted 5' to the iso-2-cytochrome c coding region of CYC7, causes a 20-fold increase in CYC7 expression. Deletion analysis of CYC7-H2 has shown that distal regions of the Ty1 element are not essential for the transcriptional activation at CYC7. In this report, we have analyzed Ty1 and CYC7 RNA from two CYC7-H2 deletion derivative genes to determine whether a direct correlation exists between transcription of Ty1 and transcription of the adjacent gene. Assuming that all Ty1 elements in the genome are transcribed equally, amounts of CYC7-H2 deletion derivative Ty1 RNA were found to be at least fivefold lower than the amount estimated for the average Ty1 element. These same Ty1 deletion derivatives caused a 20-fold increase in adjacent CYC7 expression. This finding suggests that the mechanism by which Ty1 activates adjacent gene expression does not require normal levels of Ty1 transcription. Two inversion derivatives of the CYC7-H2 Ty1 have also been analyzed. These derivatives did not produce any iso-2-cytochrome c or any normal CYC7 mRNA. Instead they were found to produce a Tyl-CYC7 fusion RNA. Consistent with our findings on CYC7-H2 Ty1 transcription, the amount of the fusion RNA was very low. In addition, the Ty1 inversion derivatives produced a new RNA that mapped to sequences upstream from the inverted Ty1 segment. Similar to Ty1 insertions that activate transcription, the new RNA was found to be transcribed away from Ty1.

scholarly journals Analysis of a mutator activity necessary for germline transposition and excision of Tc1 transposable elements in Caenorhabditis elegans.

Genetics ◽  
1988 ◽  
Vol 120 (2) ◽  
pp. 397-407
Author(s):  
I Mori ◽  
D G Moerman ◽  
R H Waterston
Keyword(s):  
Caenorhabditis Elegans ◽  
Transposable Elements ◽  
Transposable Element ◽  
Molecular Basis ◽  
Copy Number ◽  
Nematode Caenorhabditis Elegans ◽  
Transposable Element Family ◽  
Genetic Components ◽  
Mutator Activity ◽  
Low Copy Number

Abstract The Tc1 transposable element family of the nematode Caenorhabditis elegans consists primarily of 1.6-kb size elements. This uniformity of size is in contrast to P in Drosophila and Ac/Ds in maize. Germline transposition and excision of Tc1 are detectable in the Bergerac (BO) strain, but not in the commonly used Bristol (N2) strain. A previous study suggested that multiple genetic components are responsible for the germline Tc1 activity of the BO strain. To analyze further this mutator activity, we derived hybrid strains between the BO strain and the N2 strain. One of the hybrid strains exhibits a single locus of mutator activity, designated mut-4, which maps to LGI. Two additional mutators, mut-5 II and mut-6 IV, arose spontaneously in mut-4 harboring strains. This spontaneous appearance of mutator activity at new sites suggests that the mutator itself transposes. The single mutator-harboring strains with low Tc1 copy number generated in this study should be useful in investigations of the molecular basis of mutator activity. As a first step toward this goal, we examined the Tc1 elements in these low copy number strains for elements consistently co-segregating with mutator activity. Three possible candidates were identified: none was larger than 1.6 kb.

Functional analysis of chemically synthesized derivatives of the human CC chemokine CCL15/HCC-2, a high affinity CCR1 ligand

2008 ◽  
Vol 63 (1) ◽  
pp. 36-47 ◽  
Author(s):  
S.E. Escher ◽  
J. Vakili ◽  
U. Forssmann ◽  
A. Frimpong-Boateng ◽  
H. Sticht ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Cc Chemokine ◽  
High Affinity ◽  
Derivatives Of

scholarly journals THE Mu TRANSPOSABLE ELEMENTS OF MAIZE: EVIDENCE FOR TRANSPOSITION AND COPY NUMBER REGULATION DURING DEVELOPMENT

Genetics ◽  
1986 ◽  
Vol 112 (1) ◽  
pp. 107-119
Author(s):  
Mary Alleman ◽  
Michael Freeling
Keyword(s):  
Transposable Elements ◽  
Inbred Line ◽  
Germ Cells ◽  
Copy Number ◽  
First Generation ◽  
High Rate ◽  
Constant Number ◽  
Mutator Activity ◽  
Copy Number Regulation

ABSTRACT The Mu transposon of maize exists in a highly mutagenic strain called Robertson's Mutator. Plants of this strain contain 10-50 copies of the Mu element, whereas most maize strains and other plants have none. When Mutator plants are crossed to plants of the inbred line 1S2P, which does not have copies of Mu, the progeny plants have approximately the same number of Mu sequences as did their Mutator parent. Approximately one-half of these copies have segregated from their parent and one-half have arisen by transposition and are integrated into new positions in the genome. This maintenance of copy number can be accounted for by an extremely high rate of transposition of the Mu elements (10-15 transpositions per gamete per generation). When Mutator plants are self-pollinated, the progeny double their Mu copy number in the first generation, but maintain a constant number of Mu sequences with subsequent self-pollinations. Transposition of Mu and the events that lead to copy number maintenance occur very late in the development of the germ cells but before fertilization. A larger version of the Mu element transposes but is not necessary for transposition of the Mu sequences. The progeny of crosses with a Mutator plant occasionally lack Mutator activity; these strains retain copies of the Mu element, but these elements no longer transpose.

scholarly journals Identification of a regulatory transposon that controls the Mutator transposable element system in maize.

Genetics ◽  
1991 ◽  
Vol 129 (1) ◽  
pp. 261-270 ◽  
Author(s):  
P Chomet ◽  
D Lisch ◽  
K J Hardeman ◽  
V L Chandler ◽  
M Freeling
Keyword(s):  
Transposable Elements ◽  
Transposable Element ◽  
Restriction Fragment ◽  
Distinct Element ◽  
Single Locus ◽  
Inverted Repeats ◽  
Terminal Inverted Repeats ◽  
Mutator Activity ◽  
Element System ◽  
Multiple Copies

Abstract The Mutator system of maize consists of more than eight different classes of transposable elements each of which can be found in multiple copies. All Mu elements share the approximately 220-bp terminal inverted repeats, whereas each distinct element class is defined by its unique internal sequences. The regulation of instability of this system has been difficult to elucidate due to its multigenic inheritance. Here we present genetic experiments which demonstrate that there is a single locus, MuR1, which can regulate the transposition of Mu1 elements. We describe the cloning of members of a novel class of Mu elements, MuR, and demonstrate that a member of the class is the regulator of Mutator activity, MuR1. This conclusion is based on several criteria: MuR1 activity and a MuR-homologous restriction fragment cosegregate; when MuR1 undergoes a duplicative transposition, an additional MuR restriction fragment is observed, and MuR1 activity and the cosegregating MuR fragment are simultaneously lost within clonal somatic sectors. In addition, the MuR element hybridizes to transcripts in plants with Mutator activity. Our genetic experiments demonstrate that the MuR1 transposon is necessary to specify Mutator activity in our lines.

New derivatives of the Streptomyces temperate phage φC31 useful for the cloning and functional analysis of Streptomyces DNA

Gene ◽  
1985 ◽  
Vol 34 (2-3) ◽  
pp. 283-292 ◽  
Author(s):  
M.Rosario Rodicio ◽  
Celia J. Bruton ◽  
Keith F. Chater
Keyword(s):  
Functional Analysis ◽  
Temperate Phage ◽  
Derivatives Of

scholarly journals Transcriptional analysis of Ty1 deletion and inversion derivatives at CYC7.

1986 ◽  
Vol 6 (10) ◽  
pp. 3299-3311
Author(s):  
M Company ◽  
B Errede
Keyword(s):  
Cytochrome C ◽  
Transcriptional Activation ◽  
Fold Increase ◽  
Transcriptional Analysis ◽  
Insertion Mutation ◽  
Adjacent Gene ◽  
Coding Region ◽  
Deletion Derivative ◽  
And Inversion ◽  
Derivatives Of

One class of Ty insertion mutation in Saccharomyces cerevisiae activates expression of adjacent structural genes. The CYC7-H2 mutation, in which a Ty1 element is inserted 5' to the iso-2-cytochrome c coding region of CYC7, causes a 20-fold increase in CYC7 expression. Deletion analysis of CYC7-H2 has shown that distal regions of the Ty1 element are not essential for the transcriptional activation at CYC7. In this report, we have analyzed Ty1 and CYC7 RNA from two CYC7-H2 deletion derivative genes to determine whether a direct correlation exists between transcription of Ty1 and transcription of the adjacent gene. Assuming that all Ty1 elements in the genome are transcribed equally, amounts of CYC7-H2 deletion derivative Ty1 RNA were found to be at least fivefold lower than the amount estimated for the average Ty1 element. These same Ty1 deletion derivatives caused a 20-fold increase in adjacent CYC7 expression. This finding suggests that the mechanism by which Ty1 activates adjacent gene expression does not require normal levels of Ty1 transcription. Two inversion derivatives of the CYC7-H2 Ty1 have also been analyzed. These derivatives did not produce any iso-2-cytochrome c or any normal CYC7 mRNA. Instead they were found to produce a Tyl-CYC7 fusion RNA. Consistent with our findings on CYC7-H2 Ty1 transcription, the amount of the fusion RNA was very low. In addition, the Ty1 inversion derivatives produced a new RNA that mapped to sequences upstream from the inverted Ty1 segment. Similar to Ty1 insertions that activate transcription, the new RNA was found to be transcribed away from Ty1.

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