Study On The Transcription Level Of SpDJ-1 And Hsp3101-Hsp3105 In Schizosaccharomyces Pombe And Its Regulatory Pathway

It is well known that methylglyoxal (MG) metabolic disorder is related to many human diseases, such as diabetes and neurodegenerative diseases (Parkinson’s disease and Alzheimer’s disease). Recently, glyoxalasein (GL03) has been found to be responsible for conversion of MG without involvement of glutathione (GSH). The novel glyoxalase activity has been frequently observed in DJ-l/Hsp31 from various organisms, and sequence analysis revealed that DJ-1 and Hsp31 are from two different subfamilies of DJ-1/Hsp31/PfpI super family. Hsp31 is widely distributed in fungi and evolved from bacterium Hsp31, while DJ-1 is partially found in fungus and has lost in some fungus during evolution from bacterium YajL. Several GLO3 were identified, like EcHsp31 in Escherichia coli, GLX3 in Candida albicans and HsDJ-1 in Homo sapiens.A DJ-1 protein—SpDJ-1(SPAC22E 12.03c) and five Hsp31 proteins— Hsp3101 (SPCC757.03c), Hsp3102(SPAC5H 10.02c), Hsp3103(SPBC947.09), Hsp3104(SPAC11D3.13), Hsp3105(SPAC1F7.06) (for short:Hsp3101-Hsp3105) from Schizosaccharomyces prombe are known as homologous proteins of GLO3. Previous work has identified the GLO3 activity of SpDJ-1, Hsp3101 and Hsp3102, but little is known about the level of transcription and its regulation pathway of those GLO3 homologous genes found in S. prombe. Consequently, this study focused on the transcription and its regulation of SpDJ-1 and Hsp3101-Hsp3105, as well as those of SpDJ-1, Hsp3101 and Hsp3102 under the stimulation of MG It was showed that the expressions of SpDJ-1 and Hsp3101-Hsp3105 increase at transcriptional level in stationary phase, indicating that they might take effects during stationary phase. Spcl/Styl, a protein kinase, is required for transcriptional regulation of SpDJ-1 and Hsp3101-Hsp3105 genes. Our research showed that in stationary phase of Aspcl strain, no increases on mRNAs levels of those proteins were observed. Transcriptional factor Atfl is considered to be the main substrate of Spcl, those six genes studied in our work are transcription-dependent on Spcl. And our study also showed that transcription of SpDJ-1, HspS101, Hsp3102 and Hsp3105 depends on both Spcl and Atfl, while transcriptions of Hsp3103 and Hsp3104 only depend on Spcl. Furthermore, transcriptional factor Papl was not responsible for transcription of SpDJ-1 and Hsp3101-Hsp3105.Previous work showed that SpDJ-1, Hsp3101 and Hsp3102 display GLO3 activities. In this study, under the stimulation of methylglyoxal (MG), no significant increases on mRNA or protein level of those three genes were observed to answer MG stimulation, suggested that SpDJ-1, Hsp3101 and Hsp3102 show no functional abilities to responding MG stimulation, and the high-level expression of those genes during stationary phase might be involved in other unknown functions rather than degrading MG. On the other hand, compared with wild type strain under stimulation of MG, SpDJ-1, Hsp3101 and Hsp3102 from △papl strain showed no differences on mRNA levels, indicated that transcriptions of those genes do not depend on Pap1 under the stimulation of MG.Highlights and innovation points of this study include revealing the level of transcription and the regulation pathway of SpDJ-1 and Hsp3101-Hsp3105, and suggesting that the main functions of SpDJ-1, Hsp3101 and Hsp3102 might not involving in MG degradation. At last, our study gives a theoretical base on expression and regulation of GLO3 from Homo sapiens or other species.