Optimized mixed Markov models for motif identification

BACKGROUND: Identifying functional elements, such as transcriptional factor binding sites, is a fundamental step in reconstructing gene regulatory networks and remains a challenging issue, largely due to limited availability of training samples. RESULTS: We introduce a novel and flexible model, the Optimized Mixture Markov model (OMiMa), and related methods to allow adjustment of model complexity for different motifs. In comparison with other leading methods, OMiMa can incorporate more than the NNSplice's pairwise dependencies; OMiMa avoids model over-fitting better than the Permuted Variable Length Markov Model (PVLMM); and OMiMa requires smaller training samples than the Maximum Entropy Model (MEM). Testing on both simulated and actual data (regulatory cis-elements and splice sites), we found OMiMa's performance superior to the other leading methods in terms of prediction accuracy, required size of training data or computational time. Our OMiMa system, to our knowledge, is the only motif finding tool that incorporates automatic selection of the best model. OMiMa is freely available at [1]. CONCLUSION: Our optimized mixture of Markov models represents an alternative to the existing methods for modeling dependent structures within a biological motif. Our model is conceptually simple and effective, and can improve prediction accuracy and/or computational speed over other leading methods

Initiation of transcription of rDNA in rice

Three direct repeats of 320, 340 and 238 nucleotides were detected upstream to the 5' end of the 18S rRNA gene of an rDNA unit present on a 9.8 kb EcoRT fragment of the rice DNA. The primer extension analysis showed that the site of initiation of transcription is in the 1st repeat at an A, the 623rd nucleotide upstream to the 5' end of the 18S rRNA gene. Different stretches of the intergenic spacer DNA linked to the Chloramphenicol acetyl transferase gene were transcribed in the intact nuclei of rice embryos. The S1 nuclease protection analysis of the transcripts using [32P]-labelled Chloramphenicol acetyl transferase gene as the probe showed the presence of multiple promoters for rDNA transcription

Putative Termination Sites for rDNA Transcription in Rice

A clone bearing a 9.8 kb insert DNA containing the rDNA unit was identified by screening an EcoR1 library of rice DNA in $\gamma$ Charon 4 phage with $[^{32}P]$-rRNAs. The S1 nuclease mapping of the rDNA-precursor rRNA hybrids showed the presence of two transcription termini on the rDNA. They were mapped at positions 616 and 620 nucleotides downstream to the end of the 25S rRNA gene. The 18 nucleotide sequence, where the transcription terminates on the rDNA in rice and mice are homologous albeit in the reverse orientation

Putative termination sites for rDNA transcription in rice

A clone bearing a 9.8 kb insert DNA containing the rDNA unit was identified by screening an EcoR1 library of rice DNA in λ Charon 4 phage with [32P]-rRNAs. The S1 nuclease mapping of the rDNA-precursor rRNA hybrids showed the presence of two transcription termini on the rDNA. They were mapped at positions 616 and 620 nucleotides downstream to the end of the 25S rRNA gene. The 18 nucleotide sequence, where the transcription terminates on the rDNA in rice and mice are homologous albeit in the reverse orientation

A minor 9.8 kb rDNA unit of rice variety IR-20

A clone bearing a 9.8 kb EcoRI fragment of rice DNA contg. the genes for the rRNAs and the intergenic spacer was identified by screening in a rice genomic library in  Charon 4 phage with rRNAs. The 9.8 kb EcoRI DNA fragment was found to be a minor rDNA unit of rice variety IR-20. The rRNA genes and the intergenic spacer were mapped by hybridization and nucleotide sequence analyses. The DNAs in the intergenic spacer of the minor rDNA unit of 9.8 kb and the major rDNA unit of 8.9 kb cross-hybridized, showing that those regions are homologous

Interactions between Mei4, Rec114, and other proteins required for meiotic DNA double-strand break formation in Saccharomyces cerevisiae.

In most sexually reproducing organisms, meiotic recombination is initiated by DNA double-strand breaks (DSBs) formed by the Spo11 protein. In budding yeast, nine other proteins are also required for DSB formation, but roles of these proteins and interactions among them are poorly understood. We report here further studies of the behaviors of these proteins. Consistent with other studies, we find that Mei4 and Rec114 bind to chromosomes from leptonema through early pachynema. Both proteins showed only limited colocalization with the meiotic cohesin subunit Rec8, suggesting that Mei4 and Rec114 associated preferentially with chromatin loops. Rec114 localization was independent of other DSB factors, but Mei4 localization was strongly dependent on Rec114 and Mer2. Systematic deletion analysis identified protein regions important for a previously described two-hybrid interaction between Mei4 and Rec114. We also report functional characterization of a previously misannotated 5′ coding exon of REC102. Sequences encoded in this exon are essential for DSB formation and for Rec102 interaction with Rec104, Spo11, Rec114, and Mei4. Finally, we also examined genetic requirements for a set of previously described two-hybrid interactions that can be detected only when the reporter strain is induced to enter meiosis. This analysis reveals new functional dependencies for interactions among the DSB proteins. Taken together, these studies support the view that Mei4, Rec114, and Mer2 make up a functional subgroup that is distinct from other subgroups of the DSB proteins: Spo11-Ski8, Rec102-Rec104, and Mre11-Rad50-Xrs2. These studies also suggest that an essential function of Rec102 and Rec104 is to connect Mei4 and Rec114 to Spo11