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K J Newton

Publications and source records attributed to K J Newton.

At least 19 recordsLinked to original sources
PubMed2001

Nuclear gene dosage effects upon the expression of maize mitochondrial genes.

Each mitochondrion possesses a genome that encodes some of its own components. The nucleus encodes most of the mitochondrial proteins, including the polymerases and factors that regulate the expression of mitochondrial genes. Little is known about the number or location of these nuclear factors. B-A translocations were used to create dosage series for 14 different chromosome arms in maize plants with normal cytoplasm. The presence of one or more regulatory factors on a chromosome arm was indicated when variation of its dosage resulted in the alteration in the amount of a mitochondrial transcript. We used quantitative Northern analysis to assay the transcript levels of three mitochondrially encoded components of the cytochrome c oxidase complex (cox1, cox2, and cox3). Data for a nuclearly encoded component (cox5b) and for two mitochondrial genes that are unrelated to cytochrome c oxidase, ATP synthase alpha-subunit and 18S rRNA, were also determined. Two tissues, embryo and endosperm, were compared and most effects were found to be tissue specific. Significantly, the array of dosage effects upon mitochondrial genes was similar to what had been previously found for nuclear genes. These results support the concept that although mitochondrial genes are prokaryotic in origin, their regulation has been extensively integrated into the eukaryotic cell.

ATP Synthetase Complexes↗
PubMed1996

Involvement of S2 episomal sequences in the generation of NCS4 deletion mutation in maize mitochondria.

Cytoplasmic male sterility (CMS) and the abnormal-growth, nochromosomal stripe (NCS) phenotypes are cytoplasmically determined traits in plants. Mitochondrial DNA rearrangements involving short repeats appear to be responsible for the production of CMS reversions to fertility and NCS mutations. NCS4, a new mutant of maize CMS-S, exhibits both abnormal growth and male fertility. This mutant is unique because both mutations occurred within the same plant. Free S1 and S2 episomes normally found in CMS-S mitochondria have been lost from NCS4 plants. An S2 sequence has recombined aberrantly with a ribosomal protein coding region, rps3/rpl16. One end of the S2 sequence and the 5' end of the rps3/rpl16 transcription unit are absent from the NCS4 recombinant genome. Loss of mitochondrial ribosomal protein function is lethal; therefore, NCS4 plants are heteroplasmic for the rps3 deletion. Loss of S sequences from CMS-S mitochondria is not lethal and plants regain pollen function. Thus, although NCS4 plants have very abnormal plant phenotypes, they are male-fertile.

Base Sequence↗
PubMed1995

Evidence for a novel mitochondrial promoter preceding the cox2 gene of perennial teosintes.

We have characterized two promoters of the cytochrome oxidase subunit 2 (cox2) gene in Zea perennis mitochondria present in maize lines. Initiation at a site 907 bases upstream of the start codon results in the major approximately 1900 nt cox2 transcript. A sequence just upstream of this site conforms to the consensus described for maize mitochondrial promoters and its transcription is correctly initiated in a maize mitochondrial in vitro transcription extract. A second transcription initiation site (-347) is used only when the dominant allele of a nuclear gene, Mct, is present and its use results in an additional, shorter major transcript. Sequences flanking the Mct-dependent transcription initiation site, which we have termed the conditional promoter of cox2 (cpc), do not fit the maize mitochondrial promoter consensus and do not function in the maize in vitro transcription extract. The cpc region does not hybridize with mitochondrial, chloroplast or nuclear DNAs from most maize or teosinte lines. However, the cpc sequence is found in the same position upstream of the cox2 gene in Zea diploperennis mtDNA and it has striking similarity to the previously reported 'ORF of unknown origin' fused to the ATPase subunit 6 gene in maize CMS-C mitochondria. cpc appears to represent a new type of mitochondrial promoter. Further analysis of both conditional and constitutive promoters should help us to better understand the control of transcription in plant mitochondria.

Base Sequence↗
PubMed1994

R-type plasmids in mitochondria from a single source of Zea luxurians teosinte.

Two linear DNA plasmids resembling the R1 and R2 plasmids that are present in the mitochondria of several South American strains of maize were found in mitochondria from a single source of Zea luxurians collected by L. Mazoti. The Mazoti mtDNA is closely related to mtDNAs of other Z. luxurians, but mitochondria derived from the other Z. luxurians sources lack the plasmids. The larger plasmid from Mazoti mitochondria, M1, was cloned and large portions of it were sequenced. Restriction mapping and sequence comparisons showed that approximately 4.9 kb is similar to the S1 plasmid of maize and an additional 2.6 kb is related to R1 sequences integrated into the main mitochondrial genome of N cytoplasm. Therefore, the M1 plasmid appears to be very similar to the R1 plasmid. The inverted repeats at the ends of the M1 plasmid are not identical. The left end IR is similar to the S-TIRs found at the termini of the S plasmids. The right end IR more closely resembles the integrated R1 sequences, including the "variant" region of the TIR. Whereas the variant region contains 13 bp in the S-TIRs and 15 bp in an integrated version of R1, it is 16 bp long in M1. The region of M1 that has no homology to the S1 plasmid is expressed at very low levels in Mazoti and RU cytoplasms, but at much higher levels in CMS-S mitochondria, where part of it is present in the main mitochondrial genome.

Base Sequence↗
PubMed1994

Rescue of a maize mitochondrial cytochrome oxidase mutant by tissue culture.

The maize NCS6 mitochondrial mutation is a partial deletion of the cytochrome oxidase subunit 2 gene (cox2) that survives heteroplasmically in the plant. Mutant mitochondria segregate from normal mitochondria during somatic development giving rise to defective sectors on the plants, including areas of kernel abortion on the ears. Embryos from NCS6 kernels can be rescued by tissue culture. Slowly growing Type II callus derived from one of these embryos has been shown by PCR analysis to be homoplasmic for the mutation, carrying only the defective mitochondrial cox2 gene. Most of the rescued embryos were heteroplasmic for normal and mutant genes and heteroplasmy was maintained in the callus cultures. However, when suspension cultures were initiated from heteroplasmic calli, normal cells were shown to have a selective advantage. When the homoplasmic cox2 mutant callus cultures were placed on regeneration medium, plantlets did not regenerate. Heteroplasmic calli were capable of regeneration under the same conditions. These studies suggest that the functioning of mitochondrial cytochrome oxidase is not essential for growth as callus, but is required for the differentiation and development of plants.

Base Sequence↗
PubMed1994

The maize NCS2 abnormal growth mutant has a chimeric nad4-nad7 mitochondrial gene and is associated with reduced complex I function.

The molecular basis of the maternally inherited, heteroplasmic NCS2 mutant of maize was investigated. Analysis of the NCS2 mtDNA showed that it closely resembles the progenitor cmsT mitochondrial genome, except that the mutant genome contains a fused nad4-nad7 gene and is deleted for the small fourth exon of nad4. The rearrangement has occurred at a 16-bp repeat present in the third intron of the nad4 gene and in the second intron of the nad7 gene. Transcripts containing exon 4 of the nad4 gene are greatly reduced in mtRNA preparations from heteroplasmic NCS2 plants; larger transcripts are associated with the first three nad4 exons. Identical 5' ends of the nad4 transcripts have been mapped 396 and 247 bp upstream of the start codon in mtRNAs from both NCS2 and related non-NCS plants. The putative transcription termination signal of nad4 is deleted in mutant DNA, resulting in the production of the unique longer transcripts. The complex transcript pattern associated with nad7 is also altered in the mutant. Both nad4 and nad7 encode subunits of complex I (NADH dehydrogenase) of the mitochondrial electron transfer chain. Oxygen uptake experiments show that the functioning of complex I is specifically reduced in mitochondria isolated from NCS2 mutant plants.

Base Sequence↗
PubMed1991

The NCS3 mutation: genetic evidence for the expression of ribosomal protein genes in Zea mays mitochondria.

A deletion eliminating part of a transcribed region of mitochondrial DNA (mtDNA) has been found in the maize nonchromosomal stripe 3 (NCS3) mutant. This results in the specific loss of a set of three mitochondrial RNAs consisting, in normal plants, of a 4.9 kb transcript, its 1.8 kb intron and the resulting processed mRNA of approximately 2.9 kb. In the NCS3 mitochondrial genome the DNA encoding the putative promoter and 5' end of the affected RNAs is missing. This transcribed region of normal maize mtDNA has been sequenced and the intron splice junction has been determined. The 2.9 kb processed mRNA carries two overlapping open reading frames (ORFs) with predicted amino acid sequences that show similarity to two Escherichia coli ribosomal proteins, S3 (rps3) and L16 (rpl16). The presence of severe stunting and striping in NCS3 plants correlates absolutely with the molecular changes described here. This fact and the impaired ability for mitochondrial protein synthesis by NCS3 plants indicate that one or both of these reading frames are translated to functional ribosomal proteins in normal maize mitochondria.

Amino Acid Sequence↗
PubMed1991

Chloroplast Structure and Function Is Altered in the NCS2 Maize Mitochondrial Mutant.

The nonchromosomal stripe 2 (NCS2) mutant of maize (Zea mays L.) has a DNA rearrangement in the mitochondrial genome that segregates with the abnormal growth phenotype. Yet, the NCS2 characteristic phenotype includes striped sectors of pale-green tissue on the leaves. This suggests a chloroplast abnormality. To characterize the chloroplasts present in the mutant sectors, we examined the chloroplast structure by electron microscopy, chloroplast function by radiolabeled carbon dioxide fixation and fluorescence induction kinetics, and thylakoid protein composition by polyacrylamide gel electrophoresis. The data from these analyses suggest abnormal or prematurely arrested chloroplast development. Deleterious effects of the NCS2 mutant mitochondria upon the cells of the leaf include structural and functional alterations in the both the bundle sheath and mesophyll chloroplasts.

Journal Article↗
PubMed1990

Identification of a maize nuclear gene which influences the size and number of cox2 transcripts in mitochondria of perennial ++teosintes.

The involvement of nuclear genes in mitochondrial gene expression was investigated by identifying alterations in mitochondrial gene expression that occur when teosinte cytoplasms are introduced into certain maize inbred nuclear backgrounds. The cytoplasms from the teosintes Zea perennis, Zea diploperennis, and Zea luxurians were introduced into the maize A619 or W23 lines by recurrent backcrossing. Northern analysis revealed that the Z. perennis and Z. diploperennis mitochondrial cox2 transcript patterns were dependent upon the maize nuclear genotype. In a W23 nuclear background, these teosinte mitochondria have two major transcripts of 1.9 and 1.7 kb, whereas in an A619 background, they have three major transcripts of 1.9, 1.5 and 1.3 kb. No effect of nuclear background on cox2 transcripts was observed for plants possessing Z. luxurians cytoplasm. All teosinte-maize combinations possess larger, minor cox2 transcripts of 3.9, 3.3 and 3.0 kb; nuclear background has no effect on these transcripts. Immunoblot analysis showed a threefold reduction of the COXII polypeptide in Z. perennis-A619 combinations compared to Z. perennis-W23 combinations. All the major and minor transcripts posses both cox2 exons. The cox2 intron is missing from all the major transcripts and is present only in the 3.9- and 3.0-kb minor transcripts. The 1.7- and 1.3-kb transcripts are missing untranslated regions 3' to the cox2 gene; therefore at least some of the size heterogeneity is due to differential termination or downstream processing. Genetic analyses indicate that a single nuclear gene is responsible for the observed differences in the major cox2 transcripts, and that A619 carries the dominant allele.(ABSTRACT TRUNCATED AT 250 WORDS)

Blotting, Northern↗
PubMed1990

An abnormal growth mutant in maize has a defective mitochondrial cytochrome oxidase gene.

We describe a new maternally inherited maize mutation, nonchromosomal stripe 5 (NCS5), that adversely affects plant growth and yield. Mutant plants are characterized by reduced height, defective yellow striping on leaves, and aborted kernels on ears. NCS5 striped plants carry both normal and partially deleted versions of the mitochondrial cytochrome oxidase subunit 2 gene and exhibit greatly reduced levels of cox2 transcripts when compared with nonstriped control plants. Other mitochondrial genes and their mRNAs are not affected. Thus, the defective plant phenotype is correlated with a reduction in the number of functional cytochrome oxidase subunit 2 genes. The NCS5 mutant mitochondrial genome appears to have arisen by amplification of a rare homologous recombination product.

Base Sequence↗
PubMed1990

A partially deleted mitochondrial cytochrome oxidase gene in the NCS6 abnormal growth mutant of maize.

The nonchromosomal stripe (NCS) mutations of maize are a set of maternally inherited mutations that result in reduced plant growth and lowered reproductive capacity. NCS6 mutants are characterized by stunted growth, yellow stripes on leaves, and sectors of aborted kernels on ears. The heteroplasmic NCS6 striped plants carry both partially deleted and normal versions of the mitochondrial cytochrome oxidase subunit 2 (cox2) gene and have low levels of cox2 transcripts when compared with nonstriped control plants. Other tested mitochondrial genes and their transcripts appear to be unaltered in NCS6 plants. The level of COXII protein is also specifically reduced in the striped plants relative to that of control plants. In the NCS6 mutant mitochondrial genome, a portion of the first exon of the cox2 gene is deleted along with the 5' flanking region. The formation of the defective cox2 gene appears to have involved a recombination event between 31-bp repeats.

Base Sequence↗
PubMed1989

Maize nuclear background regulates the synthesis of a 22-kDa polypeptide in Zea luxurians mitochondria.

When cytoplasm of the teosinte Zea luxurians is introduced into certain maize inbred nuclear backgrounds, the pattern of protein synthesis in the teosinte mitochondria is altered. Teosinte mitochondria purified from plants possessing a maize A619 nuclear background (Z.l.-A619 plants) synthesize a novel 22-kDa polypeptide that is associated exclusively with the membrane fraction of the organelle. Mitochondria from plants possessing a W23 nuclear background do not synthesize this protein. The F(1) hybrids Z.l.-A619 x W23 and Z.l.-W23 x A619 do not synthesize the protein. However, synthesis of the polypeptide was observed in 14 of 21 individual progeny from the backcross of the F(1) hybrid Z.l.-A619 x W23 to the pollen parent A619. These data suggest that a single nuclear gene controls the synthesis of the 22-kDa protein in mitochondria, with the recessive allele of the gene allowing expression of the polypeptide. Mitochondria from the F(1) hybrid Z.l.-A619 x Mo17 synthesize the 22-kDa protein, whereas mitochondria from Z.l.-A619 x B73 do not. Data from these outcrosses demonstrate that other maize lines also possess nuclear genes capable of regulating the synthesis of the 22-kDa Zea luxurians mitochondrial protein.

Journal Article↗
PubMed1987

Altered mitochondrial gene expression in the nonchromosomal stripe 2 mutant of maize.

The genetic and molecular analyses of higher plant mitochondria can be facilitated by studying maternally-inherited mutations, such as the nonchromosomal stripe (NCS) mutants of maize, that have deleterious effects on plant growth. We have previously demonstrated a correlation between specific alterations in mitochondrial DNA and the expression of NCS phenotypes. In the present studies, the effects of the NCS2 mutation on mitochondrial gene expression are evaluated. Proteins synthesized by mitochondria isolated from NCS2 mutants and from related plants with normal growth have been compared. NCS2 mitochondria synthesize much reduced amounts of a single polypeptide. Probes corresponding to the mitochondrial DNA region altered in NCS2 hybridize to an aberrant set of transcripts in NCS2 mitochondria. Transcripts homologous to several previously characterized plant mitochondrial genes are similar in NCS2 and related non-mutant mitochondria.

Journal Article↗
PubMed1986

Mitochondrial DNA changes in abnormal growth (nonchromosomal stripe) mutants of maize.

The genetic analysis of higher plant mitochondria has been limited by a scarcity of identified mutations with known progenitors. Correspondingly, few molecular studies have been directed at types of plant mitochondrial variation other than cytoplasmic male sterility. The maternally inherited nonchromosomal stripe (NCS) mutants of maize have profound deleterious effects on plant growth and yield. We report specific alterations in mitochondrial DNA (mtDNA) for two independent, phenotypically distinct NCS mutants. NCS2 plants have a distinctive 21-kilobase Xho I mtDNA band and very reduced amounts of DNA in an 8-kilobase band that is present in the progenitor. NCS3 plants have a distinctive 20-kilobase Xho I band and a reduction in a 16-kilobase band. Our studies confirm that the affected organelle in NCS plants is the mitochondrion. Because NCS-type plants appear with a certain frequency in a particular line (WF9), this line is a potential source of additional mutations for functional and molecular analyses of maize mitochondrial genes.

Journal Article↗
PubMed1985

Mitochondrial gene expression in Cucurbitaceae: conserved and variable features.

We have examined mitochondrial DNA (mtDNA) sequence conservation, transcriptional patterns of mitochondrial genes, and mitochondrial translation products in four species of the plant family Cucurbitaceae, in which there is a seven-fold range in mitochondrial genome size. A set of conserved mtDNA sequences which we term "core" DNA is present in all cucurbit genomes examined. In watermelon, only those mtDNA restriction fragments which contain "core" DNA hybridize with mtRNA. Similar numbers of polypeptides are synthesized by isolated mitochondria from all species examined, but an extra set of mtDNA sequences is transcribed in the largest genome (muskmelon). Taken together, these results suggest that some mtRNA is untranslated in muskmelon. Cloned mitochondrial genes of known function, from maize, identified mitochondrial transcripts that varied in both size and number among cucurbit species.

Base Sequence↗