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Biomedical subjects

P Fay

Publications and source records attributed to P Fay.

At least 19 recordsLinked to original sources

Inhibition of HIV-1 replication using a mutated tRNALys-3 primer.

Cellular tRNALys-3 serves as the primer for reverse transcription of human immunodeficiency virus, type 1 (HIV-1). tRNALys-3 interacts directly with HIV-1 reverse transcriptase, is packaged into viral particles and anneals to the primer-binding site (PBS) of the HIV-1 genome to initiate reverse transcription. Therefore, the priming step of reverse transcription is a potential target for antiviral strategies. We have developed a mutant tRNALys-3 derivative with mutations in the PBS-binding region such that priming specificity was re-directed to the highly conserved TAR stem-loop region. This mutant tRNA retains high-affinity binding to HIV-1 reverse transcriptase, viral encapsidation, and is able to prime at both the targeted TAR sequence and at the viral PBS. Constitutive expression of mutant tRNA in T-cells results in marked inhibition of HIV-1 replication, as determined by measurements of viral infectivity, syncytium formation, and p24 production. Inhibition of retroviral replication through interference with the normal process of priming constitutes a new anti-retroviral approach and also provides a novel tool for dissecting molecular aspects of priming.

Base Sequence

Oxygen relations of nitrogen fixation in cyanobacteria.

The enigmatic coexistence of O2-sensitive nitrogenase and O2-evolving photosynthesis in diazotrophic cyanobacteria has fascinated researchers for over two decades. Research efforts in the past 10 years have revealed a range of O2 sensitivity of nitrogenase in different strains of cyanobacteria and a variety of adaptations for the protection of nitrogenase from damage by both atmospheric and photosynthetic sources of O2. The most complex and apparently most efficient mechanisms for the protection of nitrogenase are incorporated in the heterocysts, the N2-fixing cells of cyanobacteria. Genetic studies indicate that the controls of heterocyst development and nitrogenase synthesis are closely interrelated and that the expression of N2 fixation (nif) genes is regulated by pO2.

Cyanobacteria

Effects of long-term treatment with acetylene on nitrogen-fixing microorganisms.

Long periods of experimental incubation with acetylene led to a multifold enhancement of acetylene-reducing activity in Anabaena cylindrica, Anabaenopsis circularis, Rhodospirillum rubrum, and Azotobacter vinelandii. Rates of acetylene reduction showed a gradual increase and reached a peak after 2 to 6 h of continuous incubation under acetylene. Thereafter, enzyme activity rapidly declined. A similar enhancement of ethylene production was observed when pretreatment with acetylene was interrupted periodically by a brief exposure to ambient (or oxygen-free) atmosphere without acetylene although the decline of acetylene-reducing activity was less rapid. Pretreatment with acetylene depressed photosynthetic 14CO2 fixation and 15N2 incorporation in Anabaena cylindrica. It is concluded that assessments based on long-term experimental incubation with acetylene may grossly overestimate the actual quantities of fixed nitrogen in the field.

Acetylene

Factors influencing dark nitrogen fixation in a blue-green alga.

Nitrogen-fixing activity declines first rapidly and then more gradually when Anabaenopsis circularis is transferred from light into dark conditions. The rate and duration of dark acetylene reduction (nitrogen fixation) depend upon conditions prevailing during the preceding light period. Factors (such as light intensity, CO2 concentration, and supply of glucose), which in the light affect photosynthesis and the accumulation of reserve carbon, have a profound effect on dark nitrogen fixation. Glucose greatly promotes nitrogen fixation in the light and supports prolonged nitrogenase activity in the dark. The results suggest that heterotrophic nitrogen fixation by blue-green algae in the field may be important both under light and dark conditions.

Acetylene