RAD18 promotes DNA double-strand break repair during G1 phase through chromatin retention of 53BP1

Recruitment of RAD18 to stalled replication forks facilitates monoubiquitination of PCNA during S-phase, promoting translesion synthesis at sites of UV irradiation-induced DNA damage. In this study, we show that RAD18 is also recruited to ionizing radiation (IR)-induced sites of DNA double-strand breaks (DSBs) forming foci which are co-localized with 53BP1, NBS1, phosphorylated ATM, BRCA1 and γ-H2AX. RAD18 associates with 53BP1 and is recruited to DSB sites in a 53BP1-dependent manner specifically during G1-phase, RAD18 monoubiquitinates KBD domain of 53BP1 at lysine 1268 in vitro. A monoubiquitination-resistant 53BP1 mutant harboring a substitution at lysine 1268 is not retained efficiently at the chromatin in the vicinity of DSBs. In Rad18-null cells, retention of 53BP1 foci, efficiency of DSB repair and post-irradiation viability are impaired compared with wild-type cells. Taken together, these results suggest that RAD18 promotes 53BP1-directed DSB repair by enhancing retention of 53BP1, possibly through an interaction between RAD18 and 53BP1 and the modification of 53BP1.     

A new insect cell line from the longicorn beetle Plagionotus christophi (Coleoptera: Cerambycidae)

We have established a continuous cell line from the fat body tissue of the longicorn beetle Plagionotus christophi. The cells have been serially subcultured in MGM450 medium, and the line has been designated as PC-1. The cells were grown in suspension and comprise largely flattened spindle- or oval-shaped cells morphologically related to blood cells of longicorn beetles. The chromosome number ranged from 19 to 36, with a mode of 20 (diploid). The growth curve was determined at the 100th passage, and the population doubling time was calculated to be 3.79 d. Isozyme analysis of malic enzyme, phosphoglucose isomerase, and phosphoglucomutase revealed that PC-1 cells were enzymatically distinct from coleopteran XP-1, AnCu-35, dipteran AeAl-2, and lepidopteran GaMe-LF1 cell lines.     

Effect of juvenile hormone on the embryogenesis of a polyembryonic wasp,copidosoma floridanum,in vitro

Summary Single two-cell-stage embryos of a polyembryonic waspCopidosoma floridanum cultured in 20 μl droplets of culture medium developed to morulae at the same developmental rate as those in host eggs, but the subsequent development into polymorulae was inferior. This inferior development was markedly improved by addition of juvenile hormone (I, II, or III) or its analogues to the culture medium in a concentration-dependent manner.     

Partial purification and characterization of DNA-dependent RNA polymerases I and II from cherry salmon (Oncorhynchus masou)

1. DNA-dependent RNA polymerases I and II were purified approx 3900- and 13,000-fold, respectively, from sonicated nuclear extract of cherry salmon (Oncorhynchus masou) liver by DEAE-Sephadex, heparin-Sepharose and DNA-cellulose column chromatography. 2. The purified RNA polymerases exhibited a requirement for four kinds of ribonucleoside 5'-triphosphates, an exogeneous template and divalent cation. 3. The activities of RNA polymerases I and II were inhibited by Actinomycin D (24 micrograms/ml) but not by Rifampicin (200 micrograms/ml). 4. RNA polymerase I preferred native DNA as template, while polymerase II preferred single-stranded DNA. 5. RNA polymerase II was inhibited by a low concentration of alpha-amanitin (0.02 micrograms/ml). RNA polymerase I was also inhibited by the relatively high concentration of alpha-amanitin (IC50 = 100 micrograms/ml and IC70 = 750 micrograms/ml). 6. RNA polymerases from cherry salmon exhibited a higher activity at low temperature than from rat liver.     

Development of extracellular electric pattern aroundLepidium roots: its possible role in root growth and gravitropism

Summary Using a vibrating probe technique, four distinct electric patterns around growing cress roots were observed. The growth rate of the root with a particular one of them was apparently faster than that with the others. No direct correlation between the intensity of electric field and the root growth rate could be found. When gravistimulation was applied to the root, the electric pattern changed to be suitable for elongation of the gravitropic curvature. It is probable that change in electric pattern is related to growth of the root under a given environment.