Mycoplasma virus L2 is an enveloped DNA virus, which has a noncytocidal productive infection cycle (with maturation by budding through the cell membrane) followed by the establishment of lysogeny in each infected cell (with the viral genome integrated into the cell chromosome). The unique properties of this virus and experimental advantages of mycoplasmas are the basis of this proposal to investigate several fundamental problems in molecular virology. The proposal has three goals. First, viral genetics will be developed in the genetic code limited mycoplasmas; this will involve DNA sequencing, transcription-translation systems, temperature-sensitive mutants, and other mycoplasma virus variants. Second, the regulatory mechanisms involved in lysogeny and integration vs. noncytocidal productive infection will be determined; this will involve transcriptional control, location of replication origin and terminus, functional analysis of temperature-sensitive mutants, and analysis of the lysogenic state. Third, events in assembly and maturation of the enveloped budding L2 virus will be investigated; this will involve assembly intermediates and membrane assembly. Therefore, this proposal will utilize the experimental advantages of a prokaryotic system to ask questions in molecular virology that generally relate to animal viruses.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032442-03
Application #
3281278
Study Section
Virology Study Section (VR)
Project Start
1983-07-01
Project End
1988-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Rochester
Department
Type
Schools of Medicine
DUNS #
208469486
City
Rochester
State
NY
Country
United States
Zip Code
14627
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Haberer, K; Haberer, A I; Cadden, S P et al. (1990) Isolation and characterization of mycoplasma virus L3 temperature-sensitive mutants. Microbios 64:111-25
Dascher, C C; Poddar, S K; Maniloff, J (1990) Heat shock response in mycoplasmas, genome-limited organisms. J Bacteriol 172:1823-7
Sladek, T L; Maniloff, J (1987) Endonuclease from Acholeplasma laidlawii strain JA1 associated with in vivo restriction of DNA containing 5-methylcytosine. Isr J Med Sci 23:423-6
Poddar, S K; Maniloff, J (1987) Mapping of mycoplasma virus DNA replication origins and termini. J Virol 61:1909-12
Poddar, S K; Maniloff, J (1986) Chromosome analysis by two-dimensional fingerprinting. Gene 49:93-102
Dybvig, K; Sladek, T L; Maniloff, J (1986) Isolation of mycoplasma virus L2 insertion variants and miniviruses. J Virol 59:584-90
Sladek, T L; Nowak, J A; Maniloff, J (1986) Mycoplasma restriction: identification of a new type of restriction specificity for DNA containing 5-methylcytosine. J Bacteriol 165:219-25
Rogers, M J; Simmons, J; Walker, R T et al. (1985) Construction of the mycoplasma evolutionary tree from 5S rRNA sequence data. Proc Natl Acad Sci U S A 82:1160-4
Sladek, T L; Maniloff, J (1985) Transfection of REP- mycoplasmas with viral single-stranded DNA. J Virol 53:25-31

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