RNA editing by cytidine insertion in mitochondria of Physarum polycephalum

A corollary of the central dogma of molecular biology is that genetic information passes from DNA to RNA by the continuous synthesis of RNA on a DNA template. The demonstration of RNA editing (the specific insertion, deletion or substitution of residues in RNA to create an RNA with a sequence different from its own template) raised the possibility that in some cases not all of the genetic information for a trait residues in the DNA template. Two different types of RNA editing have been identified in mitochondria: insertional editing represented by the extensive insertion (and occasional deletion) of uridine residues in mitochondrial RNAs of the kinetoplastid protozoa and the substitutional editing represented by the cytidine to uridine substitutions in some plant mitochondria. These editing types have not been shown to be present in the same organism and may have very different mechanisms. RNA editing of both types has been observed in nonmitochondrial systems but is not as extensive and may involve still different mechanisms. Here we report the discovery of extensive insertional RNA editing in mitochondria from an organism other than a kinetoplastid protozoan. The mitochondrial RNA apparently encoding the alpha subunit of ATP synthetase in the acellular slime mould, Physarum polycephalum, is edited at 54 sites by cytidine insertion.     

Molecular determinants and guided evolution of species-specific RNA editing

Most RNA editing systems are mechanistically diverse, informationally restorative, and scattershot in eukaryotic lineages. In contrast, genetic recoding by adenosine-to-inosine RNA editing seems common in animals; usually, altering highly conserved or invariant coding positions in proteins. Here I report striking variation between species in the recoding of synaptotagmin I (sytI). Fruitflies, mosquitoes and butterflies possess shared and species-specific sytI editing sites, all within a single exon. Honeybees, beetles and roaches do not edit sytI. The editing machinery is usually directed to modify particular adenosines by information stored in intron-mediated RNA structures. Combining comparative genomics of 34 species with mutational analysis reveals that complex, multi-domain, pre-mRNA structures solely determine species-appropriate RNA editing. One of these is a previously unreported long-range pseudoknot. I show that small changes to intronic sequences, far removed from an editing site, can transfer the species specificity of editing between RNA substrates. Taken together, these data support a phylogeny of sytI gene editing spanning more than 250 million years of hexapod evolution. The results also provide models for the genesis of RNA editing sites through the stepwise addition of structural domains, or by short walks through sequence space from ancestral structures.     

Glutamate receptors: RNA editing and death of motor neurons

The aetiology of sporadic amyotrophic lateral sclerosis (ALS), a fatal paralytic disease, is largely unknown. Here we show that there is a defect in the editing of the messenger RNA encoding the GluR2 subunit of glutamate AMPA receptors in the spinal motor neurons of individuals affected by ALS. This failure to swap an arginine for a glutamine residue at a crucial site in the subunit, which occurs normally in the affected brain areas of patients with other neurodegenerative diseases, will interfere with the correct functioning of the glutamate receptors and may be a contributory cause of neuronal death in ALS patients.     

小麦抗叶锈病相关蛋白的初步分析

以小麦抗叶锈病近等基因系TcLr10和感病亲本Thatcher为植物材料.在未接种小麦叶锈菌之前提取二者叶片的蛋白质,并在接种小麦叶锈菌之后24h,48h,72h分别提取TcLr10与Thatcher叶片的蛋白质组,将提取的全部样品进行双向电泳.对所得蛋白质双向电泳图谱进行分析,结果表明,在未接种叶锈菌时TcLr10与Thatcher存在三个点的差异,其中有两个蛋白质点在TcLr10表达量明显高于感病亲本Thatcher,另一个点在TcLr10中有表达而在Thatcher中未发现;接种叶锈菌后TcLr10与Thatcher中均诱导产生一蛋白质点,但该点在Lr10中诱导产生的速度和量均明显高于感病亲本Thatcher.     

小麦麸皮木瓜蛋白酶法提取工艺研究

采用木瓜蛋白酶提取麦麸中的蛋白质,以蛋白提取率为指标,研究了温度、pH值、固液比、酶用量、提取时间对蛋白提取率的影响.用正交实验确定木瓜蛋白酶提取麦麸蛋白较适宜的条件为固液比为1∶10,提取时间为2.5 h,温度为55℃,pH值为7.0,酶与底物比为0.6%,提取率最高达到44.34%.     

DNA-protein conjugates can enter mitochondria via the protein import pathway

Mitochondria import most of their proteins and small molecules from the cytoplasm. There is some tentative evidence that they import some of their RNAs, but it is not known how nucleic acids could enter mitochondria. Here, we show that isolated yeast mitochondria can import a single-stranded or double-stranded 24-base pair piece of DNA whose 5' end is covalently linked to the C-terminus of a mitochondrial precursor protein.