In eukaryotic cells, chromatin serves as the physiological template for gene transcription, DNA replication, and repair. Chromatin assembly factor 1 (CAF-1) is the prime candidate protein to mediate assembly of newly replicated DNA into chromatin. To investigate the physiological role of CAF-1 in vivo, we used RNA interference (RNAi) to silence the 60-kDa subunit of CAF-1 (p60) in human cells. Transfection of a small interfering RNA (siRNA) directed against p60 resulted in efficient silencing of p60 expression within 24 h. This silencing led to an induction of programmed cell death in proliferating but not in quiescent human cells. Concomitantly, proliferating cells lacking p60 accumulated DNA double-strand breaks and increased levels of the phosphorylated histone H2A.X. Nuclear extracts from cells lacking p60 exhibited a 10-fold reduction of nucleosome assembly activity during DNA synthesis, which was restored upon addition of recombinant p60 protein. Nascent chromatin in cell nuclei lacking p60 showed significantly increased nuclease sensitivity, indicating chromatin assembly defects during DNA synthesis in vivo. Collectively, these data identify CAF-1 as an essential factor not only for S-phase-specific chromatin assembly but also for proliferating cell viability.

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SUMMARY In light of increased cancer prevalence and cancer-specific deaths in patients with infections, we investigated whether infections alter anti-tumor immune responses. We report that acute influenza infection of the lung promotes distal melanoma growth in the dermis and leads to accelerated cancer- specific host death. Further, we show that during influenza infection anti-melanoma CD8+ T cells are shunted from the tumor to the infection site, where they express high levels of the inhibitory receptor, PD-1. Immunotherapy to block PD-1 reverses this loss of anti- tumor CD8+ T cells from the tumor and decreases infection-induced tumor growth. Our findings show that acute non-oncogenic infection can promote cancer growth, raising concerns regarding acute viral illness sequelae. They also suggest a role for PD-1 blockade in cancer immunotherapy, and provide insight into the immune response when faced with concomitant challenges.

Temporal pattern recognition based on instantaneous spike rate coding in a simple auditory system. J Neurophysiol 90: 2484 -2493, 2003; 10.1152/jn.00259.2003. Auditory pattern recognition by the CNS is a fundamental process in acoustic communication. Because crickets communicate with stereotyped patterns of constant frequency syllables, they are established models to investigate the neuronal mechanisms of auditory pattern recognition. Here we provide evidence that for the neural processing of amplitude-modulated sounds, the instantaneous spike rate rather than the time-averaged neural activity is the appropriate coding principle by comparing both coding parameters in a thoracic interneuron (Omega neuron ON1) of the cricket (Gryllus bimaculatus) auditory system. When stimulated with different temporal sound patterns, the analysis of the instantaneous spike rate demonstrates that the neuron acts as a low-pass filter for syllable patterns. The instantaneous spike rate is low at high syllable rates, but prominent peaks in the instantaneous spike rate are generated as the syllable rate resembles that of the species-specific pattern. The occurrence and repetition rate of these peaks in the neuronal discharge are sufficient to explain temporal filtering in the cricket auditory pathway as they closely match the tuning of phonotactic behavior to different sound patterns. Thus temporal filtering or "pattern recognition" occurs at an early stage in the auditory pathway.

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