Modulation of DNA end joining by nuclear proteins

DNA double strand breaks in mammalian cells are primarily repaired by homologous recombination and non-homologous end joining (NHEJ). NHEJ may either be error-free or mutagenic with deletions or insertions at the joint. Recent studies showed that DNA ends can also be joined via microhomologous sequences flanking the break point especially when proteins responsible for NHEJ, such as Ku, are absent. Microhomology-mediated end joining (MHEJ) is always accompanied by a deletion that spans one of the two homologous sequences and the intervening sequence, if any. In this study we evaluated several factors affecting the relative contribution of MHEJ to DNA end joining using nuclear extracts and DNA substrates containing 10-bp repeats at the ends. We found that the occurrence of MHEJ is determined by the relative abundance of nuclear proteins. At low DNA/protein ratios, an error-free end-joining mechanism predominated over MHEJ. As the DNA/protein ratio increased, MHEJ became predominant. We show that the nuclear proteins that contribute to the inhibition of the error-prone MHEJ include Ku and histone H1. Treatment of extracts with flap endonuclease 1 antiserum significantly reduced MHEJ. Addition of a 17-bp intervening sequence between the microhomologous sequences significantly reduced the efficiency of MHEJ. Thus, this cell-free assay provides a platform for evaluating factors modulating end joining.     

Structure and Specificity of the Bacterial Cysteine Methyltransferase Effector NleE Suggests a Novel Substrate in Human DNA Repair Pathway

Enteropathogenic E. coli (EPEC) and related enterobacteria rely on a type III secretion system (T3SS) effector NleE to block host NF-κB signaling. NleE is a first in class, novel S-adenosyl-L-methionine (SAM)-dependent methyltransferase that methylates a zinc-coordinating cysteine in the Npl4-like Zinc Finger (NZF) domains in TAB2/3 adaptors in the NF-κB pathway, but its mechanism of action and other human substrates are unknown. Here we solve crystal structure of NleE-SAM complex, which reveals a methyltransferase fold different from those of known ones. The SAM, cradled snugly at the bottom of a deep and narrow cavity, adopts a unique conformation ready for nucleophilic attack by the methyl acceptor. The substrate NZF domain can be well docked into the cavity, and molecular dynamic simulation indicates that Cys673 in TAB2-NZF is spatially and energetically favorable for attacking the SAM. We further identify a new NleE substrate, ZRANB3, that functions in PCNA binding and remodeling of stalled replication forks at the DNA damage sites. Specific inactivation of the NZF domain in ZRANB3 by NleE offers a unique opportunity to suggest that ZRANB3-NZF domain functions in DNA repair processes other than ZRANB3 recruitment to DNA damage sites. Our analyses suggest a novel and unexpected link between EPEC infection, virulence proteins and genome integrity.     

RERG is a novel ras-related, estrogen-regulated and growth-inhibitory gene in breast cancer

Using microarray analysis, we identified a unique ras superfamily gene, termed RERG (ras-related and estrogen-regulated growth inhibitor), whose expression was decreased or lost in a significant percentage of primary human breast tumors that show a poor clinical prognosis. Importantly, high RERG expression correlated with expression of a set of genes that define a breast tumor subtype that is estrogen receptor-positive and associated with a slow rate of tumor cell proliferation and a favorable prognosis for these cancer patients. RERG mRNA expression was induced rapidly in MCF-7 cells stimulated by beta-estradiol and repressed by tamoxifen treatment. Like Ras, RERG protein exhibited intrinsic GDP/GTP binding and GTP hydrolysis activity. Unlike Ras proteins, RERG lacks a known recognition signal for COOH-terminal prenylation and was localized primarily in the cytoplasm. Expression of RERG protein in MCF-7 breast carcinoma cells resulted in a significant inhibition of both anchorage-dependent and anchorage-independent growth in vitro and inhibited tumor formation in nude mice. These features of RERG are strikingly different from most Ras superfamily GTP-binding pro-teins and suggest that the loss of RERG expression may contribute to breast tumorigenesis.     

SERPINs shelter the endowed migrants in a hostile land

Since metastatic lesions of solid tumors are the major cause of mortality in cancer patients, understanding the molecular mechanisms of metastasis is of paramount importance. Although extensive knowledge has been accumulated regarding the early steps in metastasis—starting with the departure of cancer cells from their primary sites, to their transit through the hematogenous and/or lymphatic systems, and ending with their entrance into the parenchyma of distant organs—it is difficult if not impossible to translate such knowledge into medicine due to the challenge of identifying patients with only primary tumors but otherwise pristine organs. In other words, autopsy studies indicate that a large proportion of patients already harbor dormant, undetectable micrometastases at the time of cancer diagnosis (Hensel et al, 2013 ). Accordingly, stopping tumor cell dissemination is too late for these patients. Therefore, understanding the survival and outgrowth of micrometastases may hold greater promise to combat metastatic disease.     

Reversed Hyperbolic Plasmonic Responses in Phosphorene Under Uniaxial Strain

In this paper, hyperbolic plasmonic responses of phosphorene under uniaxial strains have been explored within density functional theory. In the hyperbolic regime, plasmonic slab waveguide modes are found only along armchair direction. Then, uniaxial strains up to 10% have been applied along zigzag and armchair directions, which can significantly modify its plasmonic responses. Under appropriate strain, the signs of permittivities along two in-plane directions can be even reversed, causing switching of the propagating direction of the plasmonic modes into zigzag direction. Our investigations may give a general idea about how to control the hyperbolic plasmonic modes in phosphorene via strain.

     

Responses of soil microbial communities to prescribed burning in two paired vegetation sites in southern China

Prescribed burning is a common site preparation practice for forest plantation in southern China. However, the effects of prescribed burning on soil microbial communities are poorly understood. This study examined changes in microbial community structure, measured by phospholipid fatty acids (PLFAs), after a single prescribed burning in two paired vegetation sites in southern China. The results showed that the total amount of PLFA (totPLFA) was similar under two vegetation types in the wet season but differed among vegetation type in the dry season, and was affected significantly by burning treatment only in the wet season. Bacterial PLFA (bactPLFA) and fungal PLFA (fungPLFA) in burned plots all decreased compared to the unburned plots in both seasons (P = 0.059). Fungi appeared more sensitive to prescribed burning than bacteria. Both G⁺ bacterial PLFA and G⁻ bacterial PLFA were decreased by the burning treatment in both dry and wet seasons. Principal component analysis of PLFAs showed that the burning treatment induced a shift in soil microbial community structure. The variation in soil microbial community structure was correlated significantly to soil organic carbon, total nitrogen, available phosphorus and exchangeable potassium. Our results suggest that prescribed burning results in short-term changes in soil microbial communities but the long-term effects of prescribed burning on soil microbial community remain unknown and merit further investigation.     

Extreme learning machine classification method for lower limb movement recognition

In order to identify the lower limb movements accurately and quickly, a recognition method based on extreme learning machine (ELM) is proposed. The recognizing target set is constructed by decomposing the daily actions into different segments. To get the recognition accuracy of seven movements based on the surface electromyography, the recognition feature vector space is established by integrating short-time statistical characteristics under time domain, and locally linear embedding algorithm is used to reduce the computational complexity and improve robustness of algorithm. Compared with BP, the overall recognition accuracy for each subject in the best dimension with ELM is above 95%.