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M R Hanson

Publications and source records attributed to M R Hanson.

At least 55 records · Page 3Linked to original sources

A single nuclear gene specifies the abundance and extent of RNA editing of a plant mitochondrial transcript.

A number of cytosines are altered to be recognized as uridines in transcripts of the NADH-dehydrogenase subunit 3 (nad3) gene in the mitochondria of the higher plant Petunia hybrida. Here we show that the extent of editing for three of the edit sites, all of which change the encoded amino acid, varies between different Petunia lines. Genetic analysis indicates that a single nuclear gene is responsible for this variation. Interestingly, according to RNA blot hybridization analysis, RNA editing extent and transcript abundance are correlated. This observation is consistent with the hypothesis that RNA editing is a post-transcriptional event.

Base Sequence↗

Ribosomal protein S19 is encoded by the mitochondrial genome in Petunia hybrida.

The rps19 ribosomal protein gene, which has not been previously reported in any mitochondrial genome, was identified by sequence analysis in the mitochondrial DNA of the higher plant Petunia hybrida. According to the sequence of eight rps19 cDNAs, seven C to U conversions with respect to the genomic sequence are present in rps19 transcripts. Not all transcripts are fully edited at these seven sites. Six of the seven C to U conversions change the encoded amino acid sequence by altering four codons. The rps19 gene is located entirely within a repeat sequence which is present in three copies on the 443 kb genome. Due to intragenomic recombination across these repeats, Petunia rps19 is present in nine different genomic environments.

Amino Acid Sequence↗

Transcription of the Petunia mitochondrial CMS-associated Pcf locus in male sterile and fertility-restored lines.

Transcripts of the Petunia mitochondrial cytoplasmic male sterility (CMS)-associated S-Pcf locus, which consists of three co-transcribed genes (pcf, NADH dehydrogenase subunit 3, and ribosomal protein S12), have been characterized in reproductive tissues of CMS and fertility restored (Rf) Petunia lines by nuclease protection experiments and by RNA blot hybridization. Three 5' transcript termini have been previously described. Two 3' transcript termini and an additional S-Pcf locus transcript have now been identified. The relative abundance of the three 5' transcript termini is influenced by the presence of the nuclear Rf gene. A decrease in the abundance of the -121 5' transcript terminus relative to the -266 and -522 termini is consistently seen in Petunia lines which are restored to fertility by the Rf gene, compared to CMS Petunia lines. An additional transcript with a 5' terminus within the urf-s region of pcf is much more abundant in immature bud and anther tissue than in leaf or suspension cells. The total abundance of pcf transcripts varies greatly between plants of different nuclear backgrounds which lack the nuclear Rf allele, indicating that other nuclear genes also influence expression of the S-Pcf locus.

Autoradiography↗

Splicing of the Petunia cytochrome oxidase subunit II intron.

A comparative analysis of the plant intron-containing mitochondrial cytochrome oxidase subunit II (coxII) genes provides an indication that four conserved sequence motifs, present in exon 1 (intron-binding sequences; IBS), and complementary motifs (exon-binding sequences; EBS), present in domain I of the group II intron, may be involved in splicing of the intron. Two of these potential IBS motifs (IBS1 and IBS2) have been previously discussed. Two further potential IBS motifs (IBSa and IBSb), which occur twice within exon 1, could be involved in specification of the 5' splice site and of a 5' cryptic splice site. Nuclease-protection experiments and DNA sequence analysis of a spliced coxII cDNA have confirmed the predicted positions of the petunia coxII 5' and 3' splice sites. Evidence for the occurrence of splicing in vivo at the putative 5' cryptic splice site in petunia is provided by the detection of a nuclease-protected fragment corresponding to the size which is predicted if splicing at the proposed cryptic splice site occurs. The existence and location of a cryptic splice site, upstream of the normal coxII 5' splice site, is consistent with the proposed derivation of the cytoplasmic male sterility (CMS)-associated pcf gene from an abnormally spliced coxII transcript (Pruitt and Hanson 1989).

Base Sequence↗

A termination codon is created by RNA editing in the petunia atp9 transcript.

Analysis of the cDNA of the atp9-1 gene transcript from petunia mitochondria has revealed that ten C residues of the gene sequence are edited into U in the mRNA. Seven of these edits result in amino acid changes and one introduces a stop codon before the end of the open reading frame predicted from the gene sequence. The resulting protein is better conserved when compared to the same protein in other organisms. Comparison of the edited petunia sequence with other plant mitochondrial atp9 gene sequences idicates variation in the number and positions of edits required to obtain the same amino acids in ATP9 polypeptides of higher plants.

Amino Acid Sequence↗

The male sterility-associated pcf gene and the normal atp9-1 gene in Petunia are located on different mitochondrial DNA molecules.

A mitochondrial DNA (mtDNA) region termed the S-pcf locus has previously been correlated with cytoplasmic male sterility (CMS) in Petunia. In order to understand the relationship of the S-pcf locus to homologous sequences found elsewhere in mtDNAs of both CMS and fertile lines, the structure of the mitochondrial genome of CMS Petunia line 3688 was determined by cosmid walking. The S-pcf locus, which includes the only copies of genes for NADH dehydrogenase subunit 3 (nad3) and small ribosomal subunit protein 12 (rps12) was found to be located on a circular map of 396 kb, while a second almost identical circular map of 407 kb carries the only copies of the genes for 18S and 5S rRNA (rrn18 and rrn5), the only copy of a conserved unidentified gene (orf25), and the only known functional copy of atp9. Three different copies of a recombination repeat were found in six genomic environments, predicting sub-genomic circles of 277, 266 and 130 kb. The ratio of atp9 to S-pcf mtDNA sequences was approximately 1.5 to 1, indicating that sub-genomic molecules carrying these genes differ in abundance. Comparison of the mtDNA organization of the CMS line with that of the master circle of fertile Petunia line 3704 reveals numerous changes in order and orientation of ten different sectors.

Blotting, Southern↗

Multiple trans-splicing events are required to produce a mature nad1 transcript in a plant mitochondrion.

The mitochondrial gene encoding NADH dehydrogenase subunit 1 (nad1) in Petunia hybrida is split into five exons, a, b, c, d, and e. With the use of a complete restriction map of the 443-kb Petunia mitochondrial genome, we have cloned these exons and mapped their location. Exon a is located 130 kb away from and in the opposite orientation from exons b and c. Exon d maps 95 kb away and in the opposite orientation from exons b and c. Exons d and e are separated by 190 kb. By performing the polymerase chain reaction on Petunia cDNAs, we have shown that transcripts from these five exons are joined via a series of cis- and trans-splicing events to create a mature nad1 transcript. In addition, we have found 23 C----U RNA edit sites in Petunia nad1. RNA editing changes 19 of the amino acids predicted by the genomic sequence.

Amino Acid Sequence↗

Editing of pre-mRNAs can occur before cis- and trans-splicing in Petunia mitochondria.

Plant mitochondrial mRNAs have recently been shown to undergo editing, involving cytidine-to-uridine changes relative to the DNA sequence. We have examined the temporal relationship of editing and intron removal in coxII mRNAs in Petunia mitochondria. By using differential hybridization to probes specific for edited and unedited RNA and by sequencing of individual unspliced coxII pre-mRNA cDNAs, we found that RNA editing at any editing site can precede the splicing event. Similar results were obtained from examinations of pre-mRNA cDNAs of nad1, a gene composed of multiple exons that are both cis and trans spliced. Thus, intron removal is not required before editing can occur. The existence of editing intermediates indicates that the editing process is not strictly coincident with transcription.

Antisense Elements (Genetics)↗

Plant mitochondrial mutations and male sterility.

A characteristic of CMS mutations in plants, in contrast to the single base changes in human mitochondrial mutants (49, 140), is the presence of chimeric genes or chimeric loci; different open reading frames are joined together, or placed in proximal locations and cotranscribed with standard mitochondrial genes. Despite much progress, and the identification of several mitochondrial loci that specify CMS, the molecular basis of this defect is not understood in any plant species. Observations of altered electron transport in Petunia and toxin-mediated membrane disruption in maize plants, bacteria, and yeast expressing the maize urf13 gene product, provide clues to possible mechanisms for disruption of pollen development. Whether disruption in a particular mitochondrial function is at the root of CMS in all species, or whether defects in numerous mitochondrial activities can produce sterility, will only be revealed by further probing of physiological and biochemical defects present in CMS genotypes.

Agriculture↗

Differential Mitochondrial Electron Transport through the Cyanide-Sensitive and Cyanide-Insensitive Pathways in Isonuclear Lines of Cytoplasmic Male Sterile, Male Fertile, and Restored Petunia.

Three pairs of isonuclear lines of cytoplasmic male sterile (CMS) and fertile Petunia cells (Petunia hybrida [Hook] Vilm. and Petunia parodii L.S.M.) grown in suspension culture were examined for sensitivity to inhibitors of respiratory electron transport at time-points after transfer into fresh media. Cells from CMS lines differed from cells of fertile lines in their utilization of the cyanide-insensitive oxidase pathway. Under our culture regime, after approximately 3 days of culture cells from the CMS lines exhibited much lower cyanide-insensitive, salicylhydroxamic acid-sensitive respiration than cells from the fertile lines. This respiratory difference was shown to be specific to the mitochondrial alternative oxidase pathway by using other characteristic inhibitors of mitochondrial electron transport in experiments with isolated mitochondria. Immature anthers from CMS plants also showed lower alternative oxidase activity relative to anthers from male fertile plants, but no such difference was detected in leaf tissue, ovary or perianth tissue, or anthers collected just prior to anthesis. A cell line from a fertile plant carrying a nuclear fertility restorer gene and the CMS cytoplasm exhibited increased activity of the alternative pathway compared with the CMS lines.

Journal Article↗

Human immunodeficiency virus type 1 proficiency testing. The American Association of Blood Banks/College of American Pathologists Program.

The American Association of Blood Banks/College of American Pathologists Viral Marker Survey program added samples to evaluate participant performance with test systems for the detection of antibodies to human immunodeficiency virus type 1 in 1985. On the 80 challenges sent through April 1989, the major problem observed with enzyme immunoassay testing was unexpected reactivity on samples that did not contain anti-human immunodeficiency virus type 1. One manufacturer's testing system accounted for most of the specificity problems. The sensitivity of the enzyme immunoassay for anti-human immunodeficiency virus type 1 is close to 99%, based on the multiple reactive samples used on the 16 panels. Between 1985 and 1989, there was a 14-fold increase in participants reporting Western blot results. The introduction of a licensed system for Western blots increased the number of samples that contained antibody to human immunodeficiency virus type 1 interpreted as indeterminate. Answers to supplementary questions showed that the rate of repeatably reactive and confirmed positives on blood donors dropped to less than 0.1%.

AIDS Serodiagnosis↗

Three copies of a single recombination repeat occur on the 443 kb master circle of the Petunia hybrida 3704 mitochondrial genome.

At 443 kb, the map of Petunia hybrida line 3704 mitochondrial DNA is the largest yet produced from a dicot plant. Regions of similarity to known plant mitochondrial genes and to the chloroplast genome have been placed on a master circle. One long repeated sequence, apparently active in recombination, is present in three copies. Two copies of 6.6 kb occur in a direct orientation and are separated by 199 kb. A third truncated copy of 3.5 kb is inverted relative to the other two and is separated from the others by 99 and 145 kb. The presence of the recombination repeats predicts a multipartite molecular organization, consisting of four master circles and three subgenomic circles. Two other repeated regions were found not to be substrates for, or products of recombination. The absence of recombination at certain reiterated regions indicates that there is specificity of recombination at the recombination repeats.

Chloroplasts↗

The dizzy patient. A practical approach to management.

Dizziness can generally be divided into true vertigo and pseudovertigo (giddiness or light-headedness). The most common causes of pseudovertigo are hyperventilation, orthostatic hypotension, and multisensory deficits of older patients. Of the many types of true vertigo, only a few are caused by serious structural disorders of the brainstem, and these can usually be recognized by their temporal profile and concomitant symptoms and signs. Most cases of vertigo are caused by peripheral vestibular disorders that are self-limiting. Treatment is directed toward control of the acute autonomic symptoms and labyrinthine suppression until physiologic compensation takes place. Patients with vertigo that is prolonged, chronic, and recurrent may be helped by exercises designed to hasten or assist recovery of compensatory mechanisms.

Dizziness↗

A NADH dehydrogenase subunit gene is co-transcribed with the abnormal Petunia mitochondrial gene associated with cytoplasmic male sterility.

DNA sequence analysis 3' to the Petunia S-pcf coding region has resulted in the identification of an open reading frame similar to mammalian mitochondrial genes for subunit 3 of the NADH dehydrogenase complex (nad3). Both the abnormal fused gene S-pcf and S-nad3 fall within the mitochondrial DNA region previously shown to be associated with cytoplasmic male sterility (CMS). The S-nad3 sequence, co-transcribed with S-pcf, is present in only one copy within the Petunia CMS genome. A homologous transcribed sequence from the mitochondrial genome of a fertile Petunia line has been identified. The coding region of the two genes are identical and they share homology for at least 800 bp downstream. The genes diverge 117 bp upstream of the nad3 start codon. Transcripts of the S-pcf/S-nad3 transcripts are similar in tissues of a fertility-restored line and a CMS line.

Amino Acid Sequence↗

Cytochrome oxidase subunit II sequences in Petunia mitochondria: two intron-containing genes and an intron-less pseudogene associated with cytoplasmic male sterility.

The mitochondrial genome of Petunia hybrida contains two transcribed cytochrome oxidase subunit II (coxII) genes. The coding region of both genes is split by a 1.3 kb group II intron. Unlike coxII-1, which is similar to other sequenced plant coxII genes, the coxII-2 coding region is extended by 48 codons. The cytoplasmic male sterile (CMS) Petunia contains one coxII gene similar in structure and transcript pattern to the coxII-1 gene found in the fertile genome. Comparison of the sequenced coxII genes from the fertile mitochondrial genome with the coxII sequences present in the CMS-associated pcf gene from the CMS genome (Young and Hanson 1987) suggests that pcf is a processed pseudogene. A model for the generation of pcf is presented.

Amino Acid Sequence↗

Identification of a mitochondrial protein associated with cytoplasmic male sterility in petunia.

The petunia fused gene (pcf), which is associated with cytoplasmic male sterility (CMS), is composed of sequences derived from atp9, coxII, and an unidentified reading frame termed urfS. To determine whether the pcf gene is expressed at the protein level, we produced antibodies to synthetic peptides specified by the coxII and urfS portions of the pcf gene. Anti-COXII peptide antibodies recognized petunia COXII but no other mitochondrial proteins. Anti-URF-S peptide antibodies recognized a 20-kilodalton protein present in both cytoplasmic male sterile and fertile lines and a protein with an apparent molecular mass of 25 kilodaltons present only in cytoplasmic male sterile lines. The 25-kilodalton protein was found to be synthesized by isolated mitochondria and to fractionate into both the soluble and membrane portions of disrupted mitochondria, whereas the 20-kilodalton protein was found only in the membrane fraction. The abundance of the 25-kilodalton protein was much lower in fertile plants carrying the cytoplasmic male sterile cytoplasm and a single dominant nuclear fertility restorer gene, Rf. Thus, the pcf gene is correlated with cytoplasmic male sterility not only by its co-segregation with the phenotype in somatic hybrids, but also by the modification of its expression at the protein level through the action of a nuclear gene that confers fertility.

Amino Acid Sequence↗

An experimental study of acoustoelectric transducers with nonuniform distribution of the piezoelectric coefficient.

In this paper, an experimental test of a theoretical model published previously is presented that describes the behavior of an acoustoelectric transducer with a nonuniform distribution of the piezoelectric coefficient within its bulk. Results of this theoretical model are first reviewed. Uniform and nonuniform piezoelectric transducers were fabricated, following a procedure described herein. The receive transfer functions of the transducers were recorded experimentally, and a comparison is made with the theoretical transfer functions predicted by the model, which shows good agreement. The transmit transfer functions of the uniform and nonuniform transducers were also measured and are reported. Numerical calculations of the different transfer functions given by the theoretical model for a uniform transducer associated with different backing materials are also presented, and the results are shown to be equivalent to the results following from the Mason equivalent circuit. Comparisons with experimental results and with Mason's equivalent circuit verified the new theoretical model.

Electronics↗