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1.
Appl Environ Microbiol ; 67(7): 2982-92, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11425711

RESUMO

The inoculum size effect in the dimorphic fungus Candida albicans results from production of an extracellular quorum-sensing molecule (QSM). This molecule prevents mycelial development in both a growth morphology assay and a differentiation assay using three chemically distinct triggers for germ tube formation (GTF): L-proline, N-acetylglucosamine, and serum (either pig or fetal bovine). In all cases, the presence of QSM prevents the yeast-to-mycelium conversion, resulting in actively budding yeasts without influencing cellular growth rates. QSM exhibits general cross-reactivity within C. albicans in that supernatants from strain A72 are active on five other strains of C. albicans and vice versa. The QSM excreted by C. albicans is farnesol (C(15)H(26)O; molecular weight, 222.37). QSM is extracellular, and is produced continuously during growth and over a temperature range from 23 to 43 degrees C, in amounts roughly proportional to the CFU/milliliter. Production is not dependent on the type of carbon source nor nitrogen source or on the chemical nature of the growth medium. Both commercial mixed isomer and (E,E)-farnesol exhibited QSM activity (the ability to prevent GTF) at a level sufficient to account for all the QSM activity present in C. albicans supernatants, i.e., 50% GTF at ca. 30 to 35 microM. Nerolidol was ca. two times less active than farnesol. Neither geraniol (C(10)), geranylgeraniol (C(20)), nor farnesyl pyrophosphate had any QSM activity.


Assuntos
Candida albicans/crescimento & desenvolvimento , Candida albicans/metabolismo , Farneseno Álcool/metabolismo , Farneseno Álcool/farmacologia , Candida albicans/genética , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Meios de Cultura , Farneseno Álcool/química , Farneseno Álcool/isolamento & purificação , Cromatografia Gasosa-Espectrometria de Massas , Regulação Fúngica da Expressão Gênica , Temperatura Alta
2.
Virology ; 263(1): 254-62, 1999 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-10544099

RESUMO

Sequence analysis of the 330-kb genome of chlorella virus Paramecium bursaria chlorella virus 1 (PBCV-1) revealed an open reading frame, A237R, that encodes a protein with 34% amino acid identity to homospermidine synthase from Rhodopseudomonas viridis. Expression of the a237r gene product in Escherichia coli established that the recombinant enzyme catalyzes the NAD(+)-dependent formation of homospermidine from two molecules of putrescine. The a237r gene is expressed late in PBCV-1 infection. Both uninfected and PBCV-1-infected chlorella, as well as PBCV-1 virions, contain homospermidine, along with the more common polyamines putrescine, spermidine, and cadaverine. The total number of polyamine molecules per virion ( approximately 539) is too small to significantly neutralize the virus double-stranded DNA (>660,000 nucleotides). Consequently, the biological significance of the homospermidine synthase gene is unknown. However, the gene is widespread among the chlorella viruses. To our knowledge, this is the first report of a virus encoding an enzyme involved in polyamine biosynthesis.


Assuntos
Alquil e Aril Transferases/genética , Chlorella/virologia , Phycodnaviridae/enzimologia , Phycodnaviridae/genética , Doenças das Plantas/virologia , Alquil e Aril Transferases/química , Alquil e Aril Transferases/metabolismo , Sequência de Aminoácidos , DNA/análise , DNA Viral/análise , DNA Viral/genética , Genes Virais , Dados de Sequência Molecular , Ornitina Descarboxilase/metabolismo , Poliaminas/metabolismo , Proteínas Recombinantes/metabolismo , Espermidina/biossíntese , Vírion
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