Rocha Antonio A, Malavazi Iran, Goldman Gustavo H, Puccia Rosana
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, UNIFESP, Rua Botucatu, 862, oitavo andar, 04023-062 São Paulo, SP, Brazil.
Fungal Genet Biol. 2009 Jan;46(1):85-93. doi: 10.1016/j.fgb.2008.10.009. Epub 2008 Oct 29.
We show indirect evidences for the possible involvement of NIT2-like binding motifs in transcription modulation of the PbGP43 gene, which codes for an important antigen from the human fungal pathogen Paracoccidioides brasiliensis. This investigation was motivated by the finding of 23 NIT2-like sites within the proximal -2047 nucleotides of the PbGP43 5' intergenic region from the Pb339 isolate. They compose four clusters, two of them identical. We found four NIT2-containing probes that were positive in electrophoretic mobility shift assays and further analyzed them. PbGP43 could be modulated by nitrogen primary sources in Pb339, Pb3 and Pb18 isolates, as observed by reverse transcription (RT) real time-PCR. Gene reporter assays conducted in Aspergillus nidulans suggested that the minimal fragment responsible for nitrogen modulation lies within -480 bp of the PbGP43 gene. This is the first report on PbGP43 transcription modulation in response to nitrogen primary sources, which might help understand its regulation during infection.
我们展示了间接证据,表明类NIT2结合基序可能参与了PbGP43基因的转录调控,该基因编码来自人类真菌病原体巴西副球孢子菌的一种重要抗原。这项研究的动机是在Pb339分离株的PbGP43 5'基因间隔区近端-2047个核苷酸内发现了23个类NIT2位点。它们组成四个簇,其中两个相同。我们发现四个含NIT2的探针在电泳迁移率变动分析中呈阳性,并对其进行了进一步分析。通过逆转录(RT)实时PCR观察到,PbGP43在Pb339、Pb3和Pb18分离株中可被主要氮源调节。在构巢曲霉中进行的基因报告分析表明,负责氮调节的最小片段位于PbGP43基因的-480 bp范围内。这是关于PbGP43响应主要氮源的转录调控的首次报道,这可能有助于理解其在感染过程中的调控机制。