Characterization and comparative sequence analysis of the DNA mismatch repair MSH2 and MSH7 genes from tomato

Tam, Sheh May; Samipak, Sompid; Britt, Anne B.; Chetelat, Roger T.

doi:10.1007/s10709-009-9398-3

Cited by 10 publications

(7 citation statements)

References 55 publications

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“…Thus, both RT-PCR experiments and Genevestigator data show the same biologically relevant trend in AtMSH2 gene expression in rapidly dividing tissues. This is consistent with previous reports indicating higher levels of MSH2 activity in actively dividing cells from A. thaliana [39], Zea mays [40], and Solanum lycopersicum [45] as compared to cells in mature tissues.…”

Section: Analysis Of Atmsh2 Transcript Levelssupporting

confidence: 93%

High-Level Production of MSH2 from Arabidopsis thaliana: A DNA Mismatch Repair System Key Subunit

2010

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Biochemical and immunological information concerning DNA mismatch repair proteins from higher plants is currently limited, probably due to their low abundance in vivo. An initial analysis of AtMSH2 gene expression by quantitative real-time RT-PCR indicates that calli and seedlings contain 96.7 and 1.4 cDNA copies per ng RNA, respectively, confirming that this gene is predominantly expressed in rapidly dividing tissues. In order to obtain large quantities of AtMSH2, the protein was efficiently expressed in an Escherichia coli system. The expressed gene product has an in-frame N-terminal Trx-His(6)-S-tag. The fusion protein represents about 11% of the soluble protein from IPTG-induced E. coli cells. After a two-step purification procedure the final yield accounts for 0.7 mg/g cells. Digestion of this electrophoretically homogeneous recombinant protein with enterokinase results in an intact protein with only one extra amino acid introduced at the N-terminal end. Purified intact protein was used to induce polyclonal antibodies in rabbits. These antibodies cross-react with a 110-kDa protein from cauliflower inflorescences. Together, our data describe the transcript level, cloning, expression, purification, and polyclonal antibody preparation of AtMSH2. This work will surely be useful for carrying out plant mismatch repair assays in vitro and analyzing protein expression after the exposure of plants to various stresses.

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Section: Analysis Of Atmsh2 Transcript Levelssupporting

confidence: 93%

High-Level Production of MSH2 from Arabidopsis thaliana: A DNA Mismatch Repair System Key Subunit

2010

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“…There is an increasing demand to expand the current knowledge on the MMR system from model to crop species as such studies may have direct impact on the modern agriculture. Mutation-based breeding and introgressive hybridization have long been powerful tools in crop improvement and both these processes have been linked to the functionality of MMR in plants ( Tam et al, 2009 ). It was suggested that inactivation of MMR may help induce mutagenesis with lower associated toxicity, as lower doses of mutagens would be sufficient to obtain desired traits in an MMR deficient background ( Hoffman et al, 2004 ).…”

Section: Dna Repair Pathways In Model and Crop Plants – Nature And Efmentioning

confidence: 99%

DNA damage and repair in plants – from models to crops

2015

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The genomic integrity of every organism is constantly challenged by endogenous and exogenous DNA-damaging factors. Mutagenic agents cause reduced stability of plant genome and have a deleterious effect on development, and in the case of crop species lead to yield reduction. It is crucial for all organisms, including plants, to develop efficient mechanisms for maintenance of the genome integrity. DNA repair processes have been characterized in bacterial, fungal, and mammalian model systems. The description of these processes in plants, in contrast, was initiated relatively recently and has been focused largely on the model plant Arabidopsis thaliana. Consequently, our knowledge about DNA repair in plant genomes - particularly in the genomes of crop plants - is by far more limited. However, the relatively small size of the Arabidopsis genome, its rapid life cycle and availability of various transformation methods make this species an attractive model for the study of eukaryotic DNA repair mechanisms and mutagenesis. Moreover, abnormalities in DNA repair which proved to be lethal for animal models are tolerated in plant genomes, although sensitivity to DNA damaging agents is retained. Due to the high conservation of DNA repair processes and factors mediating them among eukaryotes, genes and proteins that have been identified in model species may serve to identify homologous sequences in other species, including crop plants, in which these mechanisms are poorly understood. Crop breeding programs have provided remarkable advances in food quality and yield over the last century. Although the human population is predicted to “peak” by 2050, further advances in yield will be required to feed this population. Breeding requires genetic diversity. The biological impact of any mutagenic agent used for the creation of genetic diversity depends on the chemical nature of the induced lesions and on the efficiency and accuracy of their repair. More recent targeted mutagenesis procedures also depend on host repair processes, with different pathways yielding different products. Enhanced understanding of DNA repair processes in plants will inform and accelerate the engineering of crop genomes via both traditional and targeted approaches.

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“…Arabidopsis and other plants encode homologs of MutS and MutL proteins found in other eukaryotic organisms, but also contain a unique MSH polypeptide (MSH7) [114,115] (Table 5B). MSH7 forms a heterodimer with MSH2 and the protein complex is designated MutSc.…”

Section: Mmr In Plantsmentioning

confidence: 99%

Protecting DNA from errors and damage: an overview of DNA repair mechanisms in plants compared to mammals

Spampinato

2016

Cell. Mol. Life Sci.

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The genome integrity of all organisms is constantly threatened by replication errors and DNA damage arising from endogenous and exogenous sources. Such base pair anomalies must be accurately repaired to prevent mutagenesis and/or lethality. Thus, it is not surprising that cells have evolved multiple and partially overlapping DNA repair pathways to correct specific types of DNA errors and lesions. Great progress in unraveling these repair mechanisms at the molecular level has been made by several talented researchers, among them Tomas Lindahl, Aziz Sancar, and Paul Modrich, all three Nobel laureates in Chemistry for 2015. Much of this knowledge comes from studies performed in bacteria, yeast, and mammals and has impacted research in plant systems. Two plant features should be mentioned. Plants differ from higher eukaryotes in that they lack a reserve germline and cannot avoid environmental stresses. Therefore, plants have evolved different strategies to sustain genome fidelity through generations and continuous exposure to genotoxic stresses. These strategies include the presence of unique or multiple paralogous genes with partially overlapping DNA repair activities. Yet, in spite (or because) of these differences, plants, especially Arabidopsis thaliana, can be used as a model organism for functional studies. Some advantages of this model system are worth mentioning: short life cycle, availability of both homozygous and heterozygous lines for many genes, plant transformation techniques, tissue culture methods and reporter systems for gene expression and function studies. Here, I provide a current understanding of DNA repair genes in plants, with a special focus on A. thaliana. It is expected that this review will be a valuable resource for future functional studies in the DNA repair field, both in plants and animals.

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Characterization and comparative sequence analysis of the DNA mismatch repair MSH2 and MSH7 genes from tomato

Cited by 10 publications

References 55 publications

High-Level Production of MSH2 from Arabidopsis thaliana: A DNA Mismatch Repair System Key Subunit

High-Level Production of MSH2 from Arabidopsis thaliana: A DNA Mismatch Repair System Key Subunit

DNA damage and repair in plants – from models to crops

Protecting DNA from errors and damage: an overview of DNA repair mechanisms in plants compared to mammals

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