Streptococcus mutans, a cariogenic agent, has a glucan-binding protein gene, gbpC, and S. criceti possesses four gbpC homologs, including dblA and dblB, as does S. sobrinus. The S. criceti dblB gene encodes a 1,717-amino-acid protein having two repetitive alanine-rich and proline-rich regions and an LPXTG motif, which is recognized by the sortase SrtA, near the C terminus. Reverse transcription-PCR analysis indicated no cotranscription of the dblA and dblB genes of S. criceti. As we could not obtain a dblB mutant of S. criceti, the dblB gene was characterized in S. mutans strain GS-5, which has genetic mutations in both gbpC and spaP genes and shows an inability to agglutinate triggered by dextran. A dextran-induced agglutination assay showed that S. mutans cells carrying dblB agglutinated in the presence of dextran. A hydrophobicity assay showed that the cells containing dblB were hydrophobic. A biofilm formation assay showed that the dblB gene was associated with biofilm formation by cells cultivated in brain heart infusion broth supplemented with glucose and maltose, but not sucrose. Nucleotide sequence analysis of the S. criceti strains studied revealed a frameshift mutation in the srtA gene encoding sortase, but intact dblA and dblB genes were found in dextran-induced agglutination-negative strains, whereas intact dblA, dblB and srtA genes were found in dextran-induced agglutination-positive strains. These results suggest the cell-surface localization of dblA and dblB gene products by SrtA and the responsibility of dblB for dextran-induced agglutination, cell-surface hydrophobicity and biofilm formation in S. criceti.

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Bacteria have a variety of resistance mechanisms for surviving bacteriophage infections. Here, we describe a novel anti-phage mechanism in Escherichia coli. Cells harboring a plasmid with the genes abpA and abpB, formerly yfjL and yfjK, blocked the propagation of bacteriophages belonging to three families: T4, T2, T7 and λ phages. Both genes were necessary for the inhibition of phage propagation, and deletion of either chromosomal gene resulted in a 20% increase of progeny compared to wild-type cells. Neither overexpression nor deficiency of AbpA and AbpB had any apparent effect on E. coli growth. We isolated seven suppressor mutants of T4 phage that grew weakly on cells overexpressing AbpA and AbpB, and found that their mutations were all located in gene 41, which encodes a replicative DNA helicase that is essential for DNA replication. Furthermore, we demonstrated that AbpA and AbpB inhibited DNA replication and late gene expression of T4 phage. Similarly, DNA replication of T7 and λ phages was also inhibited by AbpA and AbpB. These results strongly suggest that E. coli AbpA and AbpB target DNA replication of phages to block their propagation.

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The lemma and palea, which enclose the pistil, stamens, and lodicules, are the most conspicuous organs in the rice spikelet. We isolated a mutant line (ng6569) in which the lemma and palea were narrower than those of the wild type, and found that the mutant had a defect in TRIANGULAR HULL1 (TH1), which encodes a nuclear protein with an ALOG domain. Detailed morphological analysis indicated that the th1 mutation caused a reduction in the size of tubercles, which are convex structures on the surface of the lemma and palea. This reduction was more pronounced in the apical region of the lemma than in the basal region, resulting in the formation of a beak-like spikelet. By contrast, the number of tubercle rows and their spatial distribution on the lemma were not affected in the th1 mutant. Thus, the TH1 gene seems to be involved in fine-tuning the morphogenesis of the lemma and palea. In situ hybridization analysis revealed that TH1 was highly expressed in the primordia of the lemma and palea, but only weakly expressed in the primordia of the sterile lemma and rudimentary glume. We then examined the effect of th1 mutation on the lemma-like structure formed in the long sterile lemma/glume1 (g1) and extra glume1 (eg1) mutants. The result showed that the th1 mutation strongly affected the morphology of the extra lemma of eg1, but had no significant effect on the transformed lemma of g1.

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We examined genetic variation in black rats (the Rattus rattus complex) from Kandy District, Sri Lanka using mitochondrial cytochrome b (cytb, 1140 bp) and nuclear melanocortin 1 receptor (Mc1r, 954 bp) gene sequences together with database sequences. We confirmed the existence of two divergent mitochondrial lineages in Sri Lankan black rats, with genetic distance of 2.2% and estimated divergence time of 0.3 million years ago. Because one lineage is unique to the island and the other is closely related to R. rattus populations on the Indian subcontinent, two migration events of R. rattus from the subcontinent are inferred, one ancient and one recent. Mc1r analyses revealed 12 haplotypes among the Sri Lankan black rats. A median-joining network together with other available sequences separated the 12 haplotypes into two groups, one unique to the island and the other related to previously reported R. rattus sequences. Notably, most individuals possessed various combinations of both haplotype groups which had no association with the cytb clades. These results imply that old and new R. rattus lineages are now intermingled as a result of hybridization in Sri Lanka. Specimens of the lesser bandicoot rat (Bandicota bengalensis) collected from Sri Lanka (n = 24) were shown to have no genetic variability in the cytb sequence. Our results indicate that the two most abundant groups of commensal rats in Sri Lanka, black rats and lesser bandicoot rats, are the product of contrasting evolutionary histories on different timescales.

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Group A rotavirus (RVA), an etiological agent of gastroenteritis in young mammals and birds, possesses a genome of 11 double-stranded RNA segments. Although it is believed that the RVA virion contains one copy of each genomic segment and that the positive-strand RNA (+RNA) is incorporated into the core shell, the packaging mechanisms of RVA are not well understood. Here, packaging signals of RVA were searched for by analyzing genomic sequences of mammalian and avian RVA, which are considered to have evolved independently without reassortment. Assuming that packaging is mediated by direct interaction between +RNA segments via base-pairing, co-evolving complementary nucleotide sites were identified within and between genomic segments. There were two pairs of co-evolving complementary sites within the segment encoding VP7 (the VP7 segment) and one pair between the NSP2 and NSP3 segments. In the VP7 segment, the co-evolving complementary sites appeared to form stem structures in both mammalian and avian RVA, supporting their functionality. In contrast, co-evolving complementary sites between the NSP2 and NSP3 segments tended to be free from base-pairings and constituted loop structures, at least in avian RVA, suggesting that they are involved in a specific interaction between these segments as a packaging signal.

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Peptide signaling plays important roles in various developmental processes of plants. Genes encoding CLE proteins, which are processed into CLE signaling peptides, are required for maintenance of the shoot apical meristem and for vascular differentiation. FON2-LIKE CLE PROTEIN1 (FCP1), a member of the CLE gene family, negatively regulates meristem maintenance in both shoot and root apical meristems of rice (Oryza sativa). Here, we examined the role of FCP1 in leaf development. We found that overexpression of FCP1 affects various aspects of leaf development in shoots regenerated from calli, making it difficult to distinguish between the leaf blade and leaf sheath. Differentiation of tissues such as vascular bundle and sclerenchyma was strongly inhibited by FCP1 overexpression. Spatial expression patterns of developmental genes DROOPING LEAF (DL) and OsPINHEAD1 (OsPNH1) were severely affected in the FCP1-overexpressing shoots. Whereas DL was expressed in the central region of leaf primordia in control shoots, DL expression was expanded throughout the leaf primordia of the FCP1-overexpressing shoots in early developmental stages. By contrast, OsPNH1, which is expressed in provascular and developing vascular tissues in normal seedlings, was strongly repressed by FCP1 overexpression. Taken together, our results suggest that FCP1 is involved in the regulation of cell fate determination during leaf development.

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