Following acetylation, newly synthesized H3-H4 is directly transferred from the histone chaperone anti-silencing factor 1 (Asf1) to chromatin assembly factor 1 (CAF-1), another histone chaperone that is critical for the deposition of H3-H4 onto replicating DNA. However, it is unknown how CAF-1 binds and delivers H3-H4 to the DNA. Here, we show that CAF-1 binds recombinant H3-H4 with 10- to 20-fold higher affinity than H2A-H2B in vitro, and H3K56Ac increases the binding affinity of CAF-1 toward H3-H4 2-fold. These results provide a quantitative thermodynamic explanation for the specific H3-H4 histone chaperone activity of CAF-1. Surprisingly, H3-H4 exists as a dimer rather than as a canonical tetramer at mid-to-low nanomolar concentrations. A single CAF-1 molecule binds a cross-linked (H3-H4)2 tetramer, or two H3-H4 dimers that contain mutations at the (H3-H4)2 tetramerization interface. These results suggest that CAF-1 binds to two H3-H4 dimers in a manner that promotes formation of a (H3-H4)2 tetramer. Consistent with this idea, we confirm that CAF-1 synchronously binds two H3-H4 dimers derived from two different histone genes in vivo. Together, the data illustrate a clear mechanism for CAF-1-associated H3-H4 chaperone activity in the context of de novo nucleosome (re)assembly following DNA replication.

Carboxymethyl cellulose (CMC) coatings alone and in combination with gamma irradiation were tested for maintaining the storage quality and extending shelf life of pear. Matured green pears were CMC coated at levels 0.25% to 1.0% w/v and gamma irradiated at 1.5 kGy. The treated fruit including control was stored under ambient (temperature 25 ± 2 °C, RH 70%) and refrigerated (temperature 3 ± 1 °C, RH 80%) conditions. Irradiation alone at 1.5 kGy gave 8 and 4 d extension in shelf life of pear following 45 and 60 d of refrigeration, respectively. CMC coating at 1.0% w/v was effective in giving 6 and 2 d extension in shelf life of pear following 45 and 60 d of refrigeration, respectively. All combinatory treatments delayed the decaying of pear during postrefrigerated storage, but combination of 1.0% w/v CMC and 1.5 kGy irradiation proved significantly (P≤0.05) effective in maintaining the storage quality and delaying the decaying of pear. The above combinatory treatment gave an extension of 12 and 6 d in shelf life of pear during postrefrigerated storage at 25 ± 2 °C, RH 70% following 45 and 60 d of refrigeration.

Freshly harvested Red delicious apples were dipped in calcium chloride solution of varying concentrations (0.5-2.0% w/v) for 1 h prior to irradiation at dose level of 0.4 kGy. Fruits after radiation treatment were stored at 2 ± 1°C, RH 90% and evaluated at intervals of 30 days for various quality parameters. Results revealed significant (p ≤ 0.05) retention in firmness, juice yield and ascorbic acid content in samples treated with combination of calcium chloride at 2.0% w/v and gamma irradiation (0.4 kGy) during storage. Water soluble pectin was inversely correlated with firmness (r = 0.88) and was significantly (p ≤ 0.05) lower in samples subjected to combination treatment of 2.0% w/v CaCl2 and 0.4 kGy irradiation throughout the storage. The combination treatment of 2.0% CaCl2 and 0.4 kGy irradiation gave about 4.3 log reduction in yeast and mold count of apple samples. Results of the post refrigeration weight loss, firmness and overall acceptability revealed that combination treatment was helpful in extending the shelf-life of Red Delicious apples by around 20-25 days at 17 ± 2°C, RH 75% following 90 days of refrigeration.

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