Two Cellular Protein Kinases, DNA-PK and PKA, Phosphorylate the Adenoviral L4-33K Protein and Have Opposite Effects on L1 Alternative RNA Splicing

Accumulation of the complex set of alternatively processed mRNA from the adenovirus major late transcription unit (MLTU) is subjected to a temporal regulation involving both changes in poly (A) site choice and alternative 3′ splice site usage. We have previously shown that the adenovirus L4-33K protein functions as an alternative splicing factor involved in activating the shift from L1-52,55K to L1-IIIa mRNA. Here we show that L4-33K specifically associates with the catalytic subunit of the DNA-dependent protein kinase (DNA-PK) in uninfected and adenovirus-infected nuclear extracts. Further, we show that L4-33K is highly phosphorylated by DNA-PK in vitro in a double stranded DNA-independent manner. Importantly, DNA-PK deficient cells show an enhanced production of the L1-IIIa mRNA suggesting an inhibitory role of DNA-PK on the temporal switch in L1 alternative RNA splicing. Moreover, we show that L4-33K also is phosphorylated by protein kinase A (PKA), and that PKA has an enhancer effect on L4-33K-stimulated L1-IIIa splicing. Hence, we demonstrate that these kinases have opposite effects on L4-33K function; DNA-PK as an inhibitor and PKA as an activator of L1-IIIa mRNA splicing. Taken together, this is the first report identifying protein kinases that phosphorylate L4-33K and to suggest novel regulatory roles for DNA-PK and PKA in adenovirus alternative RNA splicing

Are changes in behavior of fast-growing broilers with slight gait impairment (GS0-2) related to pain?

Impaired walking ability in terms of slight or definite defects is more common in broiler production than lameness that obviously hinders movement, but it has received limited scientific attention. This study aimed to compare behavior of conventional broilers with impaired walking ability (assessed as gait score (GS) 2) with those walking normally (GS0) and those with only a slight gait defect (GS1). Behavior in the home environment was registered, and an analgesic intervention to quantify changes in time budgets indicating pain relief was applied. The study included 192 Ross 308 broilers. On day 27 of age, the birds were distributed as evenly as possible into birds of GS0 and GS2 of each sex based on obtained gait score. Following this, each experimental bird was housed with 3 companion birds. On days 30 and 32 of age, the behavior in the home pens was recorded. All experimental birds were injected with the NSAID carprofen on one of the 2 d and saline on the other. The statistical analyses used the GS scored on the day of recording as explanatory factor. Compared to GS0 birds, GS2 birds tended to be more inactive (mean (CI): 4,193 (3,971–4416) vs. 4,005 (3,753–4,257) s; P = 0.074), spent more time sitting while feeding (306 (266–353) vs. 213 (180–251) s; P = 0.026), were less likely to perch (probability: 0.78 (0.69–0.85) vs. 0.91 (0.85–0.95); P = 0.012), and spent less time performing comfort behavior (749 (689–814) s vs. 875 (792–967) s; P = 0.043). Compared to GS1 birds, GS2 birds spent more time inactive (GS1: 4,022 (3,818–4225) s; P = 0.027), less time foraging (289 (253–329) vs. 347 (309–388) s; P = 0.047), and were less likely to perch (GS1: 0.90 (0.86–0.93); P = 0.001). For some of these behavioral variables, administration of carprofen led to behavioral changes across the GSs, which may suggest that the behavioral expression of the broilers was limited by pain. These findings are of relevance to animal welfare, but the underlying causes are still not fully clarified

Characterization and in vitro probiotic assessment of potential indigenous Bacillus strains isolated from soil rhizosphere

BACKGROUND: Probiotics mainly Bacillus species can be advantageous to the host by promoting its intestinal balance. Attempts were made to isolate and identify Bacillus strains from rhizosphere environment. METHODS: The in vitro probiotic criteria were used for screening and characterizing potential Bacillus probiotics. Morphological, physiological and biochemical characteristics as well as 16S rRNA gene sequence analysis were utilized for identification of the isolates. Seven isolates were chosen based on withstanding to acidic condition (pH 2.5) and various bile salt concentrations (1-4%(w/v)). RESULTS: Isolates found to have the least antimicrobial activity against Listeria monocytogenes PTCC 1163, Staphylococcus aureus ATCC 1912 and Bacillus cereus PTCC 1015; however, no activity against Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 25922 was observed. The Bacillus Isolates showed different variation in auto-aggregation features and adhesion to hydrocarbons ranging from 60% to 90% and 10% to 60%, respectively. Excluding isolate 14 that exhibited resistance to penicillin and ampicillin, all the other Bacillus strains were sensitive to the tested antibiotics. All isolates showed relatively low cytotoxic effect on HepG2 cell line except strains 12 and 14. CONCLUSION: Taking together, among the investigated Bacillus isolates, strains 17 and S10 found to be the most promising candidates to fulfill in vitro probiotic specifications