Tyrosine 842 in the activation loop is required for full transformation by the oncogenic mutant FLT3-ITD

The type III receptor tyrosine kinase FLT3 is frequently mutated in acute myeloid leukemia. Oncogenic FLT3 mutants display constitutive activity leading to aberrant cell proliferation and survival. Phosphorylation on several critical tyrosine residues is known to be essential for FLT3 signaling. Among these tyrosine residues, Y842 is located in the so-called activation loop. The position of this tyrosine residue is well conserved in all receptor tyrosine kinases. It has been reported that phosphorylation of the activation loop tyrosine is critical for catalytic activity for some but not all receptor tyrosine kinases. The role of Y842 residue in FLT3 signaling has not yet been studied. In this report, we show that Y842 is not important for FLT3 activation or ubiquitination but plays a critical role in regulating signaling downstream of the receptor as well as controlling receptor stability. We found that mutation of Y842 in the FLT3-ITD oncogenic mutant background reduced cell viability and increased apoptosis. Furthermore, the introduction of the Y842 mutation in the FLT3-ITD background led to a dramatic reduction in in vitro colony forming capacity. Additionally, mice injected with cells expressing FLT3-ITD/Y842F displayed a significant delay in tumor formation, compared to FLT3-ITD expressing cells. Microarray analysis comparing gene expression regulated by FLT3-ITD versus FLT3-ITD/Y842F demonstrated that mutation of Y842 causes suppression of anti-apoptotic genes. Furthermore, we showed that cells expressing FLT3-ITD/Y842F display impaired activity of the RAS/ERK pathway due to reduced interaction between FLT3 and SHP2 leading to reduced SHP2 activation. Thus, we suggest that Y842 is critical for FLT3-mediated RAS/ERK signaling and cellular transformation.     

Stepwise replication identifies a low-producing lymphotoxin-alpha allele as a major risk factor for early-onset leprosy

Host genetics has an important role in leprosy, and variants in the shared promoter region of PARK2 and PACRG were the first major susceptibility factors identified by positional cloning. Here we report the linkage disequilibrium mapping of the second linkage peak of our previous genome-wide scan, located close to the HLA complex. In both a Vietnamese familial sample and an Indian case-control sample, the low-producing lymphotoxin-alpha (LTA)+80 A allele was significantly associated with an increase in leprosy risk (P = 0.007 and P = 0.01, respectively). Analysis of an additional case-control sample from Brazil and an additional familial sample from Vietnam showed that the LTA+80 effect was much stronger in young individuals. In the combined sample of 298 Vietnamese familial trios, the odds ratio of leprosy for LTA+80 AA/AC versus CC subjects was 2.11 (P = 0.000024), which increased to 5.63 (P = 0.0000004) in the subsample of 121 trios of affected individuals diagnosed before 16 years of age. In addition to identifying LTA as a major gene associated with early-onset leprosy, our study highlights the critical role of case- and population-specific factors in the dissection of susceptibility variants in complex diseases.     

Genes for the major protein antigens of the leprosy parasite Mycobacterium leprae

Leprosy, a chronic infectious disease afflicting between 10 and 15 million people, is caused by the obligate intracellular parasite Mycobacterium leprae. Although M. leprae was the first identified bacterial pathogen of man, basic biochemical, immunological, diagnostic and therapeutic investigations have been severely limited because it remains one of the few human pathogens that have not been cultured in vitro. An M. leprae recombinant DNA expression library was constructed to provide a source of genes encoding proteins relevant for such studies. Monoclonal antibodies directed against M. leprae specific antigens have been used to isolate the genes encoding the five most immunogenic protein antigens of the leprosy bacillus. We report here that M. leprae specific epitopes recognized by all of 13 monoclonal antibodies tested were produced by recombinant phage in Escherichia coli.     

Comparison of the production rates of bacteria in the rumen estimated by using labelled live and formaldehyde treated mixed bacterial cells

Summary The bacteria production rates in the rumen have been estimated by injecting¹⁴C- and³⁵S-labelled mixed rumen bacteria, either live or killed by treatment with formaldehyde, into the rumen and applying isotope dilution technique. The rate of bacteria production when estimated by using either live- or dead-(protected-)labelled bacterial cells were comparable.     

Optimal policy for controlling two-server queueing systems with jockeying

This paper studies the optimal policy for joint control of admission, routing, service, and jockeying in a queueing system consisting of two exponential servers in parallel.Jobs arrive according to a Poisson process.Upon each arrival, an admission/routing decision is made, and the accepted job is routed to one of the two servers with each being associated with a queue.After each service completion, the servers have an option of serving a job from its own queue, serving a jockeying job from another queue, or staying idle.The system performance is inclusive of the revenues from accepted jobs, the costs of holding jobs in queues, the service costs and the job jockeying costs.To maximize the total expected discounted return, we formulate a Markov decision process(MDP) model for this system.The value iteration method is employed to characterize the optimal policy as a hedging point policy.Numerical studies verify the structure of the hedging point policy which is convenient for implementing control actions in practice.     

Isolation and fusion studies on protoplasts from pollen tetrads

Summary different enzymes were tested for isolation of intact protoplasts from pollen tetrads. About 80% isolation was achieved from pollen tetrads of Cajanus cajan and Zea mays and about 60% from Luffa cylindrica and Lycopersicon esculentum after 4 h of treatment with 5% cellulase. When these mononucleate protoplasts were incubated in presence of 0.05 M CaCl₂ in 0.3 M glucose at pH 10.5, 70–80% fusion was achieved. Fusion was rare in sodium nitrate solutions.This work was supported by a grant from the Indian Council of Agricultural Research, New Delhi (No. 1-32/73, Ed. I) in the form of a Senior Fellowship awarded to P. C. D.Work carried out at Molecular Cytogenetics Research Unit, Dept. of Genetics and Plant Breeding, Banaras Hindu University, Varanasi, India.