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Biomedical subjects

H Saedler

Publications and source records attributed to H Saedler.

At least 91 records · Page 5Linked to original sources
PubMed1989

Transpositional behavior of the maize En/Spm element in transgenic tobacco.

The transposable element Enhancer (En/Spm) of Zea mays encodes the functions necessary for transposition of members of the En-I family of elements. En constructs were introduced into tobacco and transposition was demonstrated by cloning and DNA sequence analysis. Transcriptional analysis of En transgenic plants revealed 2.5 and 6 kb En homologous transcripts similar in size to those found in maize, in addition to aberrant splice products. Progeny analysis of a primary transformant indicates En transpositional activity after meiosis. In these progeny somatic variegation, at the NPT II gene, was demonstrated to be caused by En excision. Two re-insertions of En were analyzed and found to reside in unique copy DNA while another was found in middle repetitive DNA.

Journal Article↗
PubMed1989

The amyloplast-targeting transit peptide of the waxy protein of maize also mediates protein transport in vitro into chloroplasts.

The transit peptide of the waxy protein of maize which in the maize plant targets this protein only into amyloplasts was used for in vitro protein transport experiments with isolated amyloplasts from maize and chloroplasts from maize, pea and potato. In the presence of both intact and disrupted amyloplasts an artificial preprotein (TP30), consisting of the waxy transit peptide plus the first 34 amino acids of the mature waxy protein fused in-frame to the beta-glucuronidase of Escherichia coli, is processed to the size expected when the transit peptide is cleaved off. The chloroplasts studied show in vitro import and correct processing of both TP30 and the authentic waxy protein, but not of the beta-glucuronidase without the waxy transit peptide. The in vitro import of TP30 into chloroplasts is almost as efficient as that of the precursor of the small subunit of pea ribulose-1,5-bisphosphate carboxylase, a nuclear-encoded chloroplast protein, whereas the waxy protein accumulates to a lesser extent in the chloroplasts. Since the amino-terminal transit peptides of TP30 and the waxy precursor are the same, this difference must be due to the mature part of the waxy protein. One possible explanation is the observed instability of the waxy protein in the presence of chloroplasts.

Amino Acid Sequence↗
PubMed1989

The maize En-1/Spm element transposes in potato.

The maize transposable element En-1 has been introduced into a diploid potato line via transformation with Agrobacterium tumefaciens. The element is transcriptionally active in potato. Numerous En specific RNAs are observed, including a 6 kb transcript characteristic of an active En-1 element in maize. In contrast to maize, where the 6.0 kb transcript is hardly detectable, this transcript is very abundant in the transgenic potato. Transposition of En-1 in the potato clone was analysed by Southern blot hybridization and confirmed by molecular isolation of En-1 excision and integration events. Sequence data of excision and integration sites revealed footprints and target site duplications similar to the ones described for En-1 in maize.

Blotting, Southern↗
PubMed1989

Multiple genes are transcribed in Hordeum vulgare and Zea mays that carry the DNA binding domain of the myb oncoproteins.

cDNA clones were isolated from tissue specific cDNA libraries of barley and maize using as a probe the cDNA of the maize gene C1, a regulator of anthocyanin gene expression. C1-related homology for all of the four cDNAs characterized by sequence analysis is restricted to the N-terminal 120 amino acids of the putative proteins. This region shows striking homology to the N-proximal domain of the myb oncoproteins from vertebrates and invertebrates. Within the myb proto-oncogene family this part of the respective gene products functions as a DNA binding domain. Acidic domains are present in the C-proximal protein segments. Conservation of these sequences, together with the genetically defined regulator function of the C1 gene product, suggest that myb-related plant genes code for trans-acting factors which regulate gene expression in a given biosynthetic pathway.

Amino Acid Sequence↗
PubMed1989

The bz-rcy allele of the Cy transposable element system of Zea mays contains a Mu-like element insertion.

The receptive component of the Cy transposable element system (rcy:Mu7) at the Bz locus of Zea mays L. is 2.2 kb and has long terminal inverted repeats. The insertion is flanked by a 9 bp duplication. In the presence of an autonomous Cy element in the genome, rcy:Mu7 is excised from bz-rcy in a manner consistent with a model suggested previously. The termini of rcy:Mu7 have 85% sequence similarity with the Mu1 element of Z. mays. This is consistent with the observation that Mu1 can behave genetically like a receptive component of the Cy system.

Alleles↗
PubMed1988

TnpA product encoded by the transposable element En-1 of Zea mays is a DNA binding protein.

TnpA protein, the function encoded by the most abundant transcript of En-1 was expressed in Escherichia coli. DNA binding experiments with partially purified tnpA protein revealed that it binds to the subterminal repetitive region of En-1. TnpA protein recognizes a 12-bp-long sequence motif which is reiterated several times at the termini of En-1. Binding is reduced if the cytosine residues of CG dinucleotides and CNG trinucleotides within the motif are methylated. These data suggest a model in which the product of tnpA serves as a regulator of element activity.

Base Sequence↗
PubMed1988

The defective En-I102 element encodes a product reducing the mutability of the En/Spm transposable element system of Zea mays.

Genetic and molecular analysis has revealed a specific En-element of deletion derivative (En-I102) which reduces En/Spm-induced mutability. In the presence of En-I102 the excision frequency of both the autonomous En-1 element and the inhibitor element Spm-I5719A is reduced and excision occurs later in development. The 3697 bp long En-I102 element is derived from En-1 by an internal deletion of 4590 bp removing nucleotides 1862-6451. The promoter at the left end and sequences required for polyadenylation are retained in En-I102. It is transcribed to yield predominantly a 1.8 kb poly(A) RNA. cDNA analysis of this transcript indicated that it contains the coding capacity for a 386 amino acid polypeptide. This polypeptide shares homology with En/Spm encoded functions and we suggest that it interferes with transposition at the protein level.

Alleles↗
PubMed1988

Transposon-induced alterations in the promoter region affect transcription of the chalcone synthase gene of Antirrhinum majus.

Four solid-colour revertants were isolated from the highly variegated niv-53::Tam1 mutant, in which the transposable element Tam1 is integrated in the promoter region of the chalcone synthase (chs) gene. DNA sequence analysis revealed that in all four lines the Tam1 element was deleted together with flanking nucleotides of the chs promoter. In one case the TATA box of the chs gene was removed resulting in extremely low expression of the gene, and initiation of transcription occurring at a new position. The other three deletions defined a sequence motif (TAC-CAT) which is apparently required for maximal gene expression. Thus transposable elements seem to be useful for probing gene structure, in this case the signal structure in the promoter region, by virtue of imprecise excision.

Acyltransferases↗
PubMed1988

Plant transposable elements: their role in evolution.

Transposable elements (TE) are natural constituents of plant genomes. However, their presence only becomes apparent if they become dislodged from their resident positions in the genome and transpose into another gene, thereby inducing a mutation. Such TE-induced mutations are somatically unstable because they revert to wild type and hence reconstitute the expression of the mutated gene. The frequent somatic excision of the TE results in a variegated phenotype. Since this instability is inherited in a Mendelian manner the variegated phenotype is nuclear determined. By this criterion TE have been shown to occur in more than 30 species belonging to different families and genera. Many questions arise when dealing with TE: their structure and functions, and the biological significance of the activity of elements in the differentiation of a normal plant or in the evolution of plant genes.

Biological Evolution↗
PubMed1988

A genomic DNA segment from Petunia hybrida leads to increased transformation frequencies and simple integration patterns.

A 2-kilobase (kb) genomic fragment was selected from Petunia hybrida that increased transformation efficiencies by at least a factor of 20 after direct DNA transfer to petunia and tobacco protoplasts when supercoiled plasmid DNA was used. Because of this effect this fragment was named transformation booster sequence (TBS). Increased transformation frequencies were observed for plasmids that contained either the 2-kb fragment in dimeric or monomeric form or an internal 1.1-kb fragment of TBS. Analysis of transformants revealed that preferentially one copy of foreign DNA is integrated. Thus, TBS improves the poor transformation frequencies of direct gene transfer using circular plasmids, while it conserves the simple integration pattern that is important for practical applications. Possible mechanisms of TBS action are discussed.

Base Sequence↗
PubMed1987

Cin4, an insert altering the structure of the A1 gene in Zea mays, exhibits properties of nonviral retrotransposons.

A wild-type allele of the A1 gene of Zea mays contains a 1.1-kb-long insert termed Cin4-1, which alters the structure of the transcription unit compared to other A1 alleles. The Cin4-1 element is a member of a family of elements occurring in 50-100 copies in the maize genome. Genomic cloning and sequence analysis of several family members and their flanking regions allowed classification of Cin4 as a nonviral retrotransposon. Individual Cin4 elements terminate in an oligo(A) track of variable size (6-11 residues) at their 3'-end. The 5'-ends of family members are heterogeneously truncated with respect to the longest Cin4 element. Cin4 elements are flanked by small direct duplications, the size of which varies between 3 and 16 bp. On the basis of a comparison of the target sequence and the sequence of Cin4 we suggest and discuss a model of the mechanism of Cin4 integration via in situ cDNA synthesis on an RNA template. The longest Cin4 element analysed so far has two non-overlapping open reading frames (ORFs) comprising 2793 nucleotides (ORF1) and 3489 nucleotides (ORF2). The putative 1163 amino acid long Cin4 protein derived from the sequence of ORF2 has the capacity to encode a reverse transcriptase-like protein and a DNA-binding domain. The conservation pattern of these two domains and the overall organisation of Cin4 is similar to that detected in nonviral retrotransposons in animals. The origin and function of Cin4 are discussed.

Journal Article↗
PubMed1987

The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators.

The structure of the wild-type c1 locus of Zea mays was determined by sequence analysis of one genomic and two cDNA clones. The coding region is composed of three exons (150 bp, 129 bp and one, at least 720 bp) and two small introns (88 bp and 145 bp). Transcription of the mRNAs corresponding to the two cDNA clones cLC6 (1.1 kb) and cLC28 (2.1 kb) starts from the same promoter. Both cDNAs are identical except that cLC28 extends further at its 3' end. A putative protein, 273 amino acids in length was deduced from the sequence of both transcripts. It contains two domains, one basic and the other acidic and might function as a transcriptional activator. The basic domain of this c1-encoded protein shows 40% sequence homology to the protein products of animal myb proto-oncogenes.

Amino Acid Sequence↗
PubMed1987

Influence of transposable elements on the structure and function of the A1 gene of Zea mays.

The structure of the A1 gene of Zea mays was determined by sequencing cDNA and genomic clones. The gene is composed of four exons and three short introns. The 40.1-kd A1 protein is an NADPH-dependent reductase. Germinal derivatives of the mutable a1-m1 allele with either recessive or wild-type phenotype have been isolated. Sequence analysis of these revertant alleles indicates that frame-shift mutations abolish A1 gene function, whereas one additional amino acid within the protein sequence still allows wild-type gene expression. The presence of a second, promoter-like structure, upstream of the functional A1 gene promoter is discussed with respect to its possible involvement in differential expression of the A1 gene. The structure of the a1-m2 8004, 3456 and 4412 alleles, featuring distinguishable phenotypes in the presence of Spm(En), was also determined. In all alleles the 1080-bp-long inhibitor (I) element is located 15 bp upstream of the CAAT box of the A1 gene promoter. The unusual response of a1-m2 alleles to trans-active signals of the Spm(En) element is discussed with respect to the position of the I inserts. Also presented are data on the structure and insertion sites of transposable elements determined by cloning and sequencing of the mutable a1 alleles a1-mpapu, a1-mr 102 and a1-ml.

Alcohol Oxidoreductases↗
PubMed1987

Molecular analysis of paramutant plants of Antirrhinum majus and the involvement of transposable elements.

Paramutation is observed when the Antirrhinum majus lines 44 and 53 are crossed. These two lines both have insertions at the nivea locus, which encodes chalcone synthase (chs). The allele niv-53 carries the transposable element Tam1 in the promoter region of the chs gene; niv-44 carries the element Tam2 within the gene. The Tam1 element has previously been extensively characterised. Here the Tam2 element is further characterised, and the arrangement of the nivea locus in paramutant plants is analysed. The complete sequence of Tam2, and that of a partial cDNA complementary to it, have been determined. The cDNA is probably transcribed from a different copy of Tam2 from that present at the nivea locus, and does not encode a functional protein. Genomic Southerns of F1 plants from the 53/44 cross show that no major rearrangements are consistently associated with paramutation at the nivea locus of A. majus. The isolation from a paramutant plant arising from a 53/44 cross of an allele (niv-4432) resulting from the excision of Tam2 is reported. The excision of Tam2 resulted in a 32 bp deletion of chs gene sequences. Plants homozygous for the new niv-4432 allele have white flowers and are still paramutagenic, demonstrating that Tam2 need not be present at the nivea locus for paramutation to occur. Different interactions between Tam1 and Tam2 are discussed, and a possible model for paramutation is presented.

Antirrhinum↗
PubMed1986

Molecular cloning of the c locus of Zea mays: a locus regulating the anthocyanin pathway.

The c locus of Zea mays, involved in the regulation of anthocyanin biosynthesis, has been cloned by transposon tagging. A clone (# 18En) containing a full size En1 element was initially isolated from the En element-induced mutable allele c-m668655. Sequences of clone # 18En flanking the En1 element were used to clone other c mutants, whose structure was predicted genetically. Clone #23En (isolated from c-m668613) contained a full size En1 element, clone #3Ds (isolated from c-m2) a Ds element and clone # 5 (isolated from c+) had no element on the cloned fragment. From these data we conclude that the clones obtained contain at least part of the c locus. Preliminary data on transcript analysis using a 1-kb DNA fragment from wild-type clone # 5 showed that at least three transcripts are encoded by that part of the locus, indicating that c is a complex locus.

Alleles↗