Delivery of HSF1(+) protein using HIV-1 TAT protein transduction domain.
HSF1 is the major transcription factor of HSPs (heat shock proteins) in response to various stresses. Wild type HSF1 (heat shock transcriptional factor 1) is normally inactive, while a constitutively active form of HSF1 (HSF1(+)) can activate downstream HSP expression in the absence of stresses. Here we generated the eukaryotic vectors that expresses HSF1(+) fusion proteins, and found that HSF1(+)-TAT fusion protein was expressed and activated HSP expression. TAT, as a trans-acting factor of HIV-1, has been demonstrated to deliver functional cargo protein into living cells. HSF1(+)-TAT fusion protein was expressed in E. coli, purified, incubated with A549 cells for 8 h, Western blot analysis and luciferase reporter assay showed that HSF1(+) fusion protein was delivered into A549 cells successfully, and the accumulation of HSF1(+)-TAT fusion protein in A549 cells up-regulated HSP70 expression.
Hou
Yonghui
Y
Guangzhou Institute of Biomedicine and Health (GIBH), Chinese Academy of Sciences, 510663 Guangzhou, China.
Zou
Jiangying
J
D000704:Analysis of Variance; D045744:Cell Line, Tumor; D002470:Cell Survival; D003001:Cloning, Molecular / Q000379:methods; D019610:Cytoprotection; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D000076249:Heat Shock Transcription Factors; D006358:Hot Temperature; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D013312:Stress, Physiological; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D000076249:Heat Shock Transcription Factors; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D054322:tat Gene Products, Human Immunodeficiency Virus