Computational analysis of the human HSPH/HSPA/DNAJ family and cloning of a human HSPH/HSPA/DNAJ expression library.
In this manuscript, we describe the generation of a gene library for the expression of HSP110/HSPH, HSP70/HSPA and HSP40/DNAJ members. First, the heat shock protein (HSP) genes were collected from the gene databases and the gene families were analyzed for expression patterns, heat inducibility, subcellular localization, and protein homology using several bioinformatics approaches. These results can be used as a working draft model until data are confirmed by experimental approaches. In addition, we describe the generation of a HSPA/DNAJ overexpression library and tested the effect of different fusion tags on HSPA and DNAJ members using different techniques for measuring chaperone activity. These results show that we have cloned a high-quality heat shock protein expression library containing most members from the HSPH, HSPA, DNAJA and DNAJB families which will be useful for the chaperone community to unravel the function of the highly diverse family of human molecular chaperones.
Hageman
Jurre
J
Section of Radiation and Stress Cell Biology, Department of Cell Biology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands. h.h.kampinga@med.umcg.nl
Kampinga
Harm H
HH
D003001:Cloning, Molecular; D019295:Computational Biology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics; D050957:HSP110 Heat-Shock Proteins / Q000235:genetics*; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D018869:Heat-Shock Response; D006801:Humans; D005810:Multigene Family / Q000235:genetics*; D009928:Organ Specificity; D010802:Phylogeny; D021381:Protein Transport; D015203:Reproducibility of Results; D040681:Structural Homology, Protein; D013347:Subcellular Fractions / Q000378:metabolism; D014158:Transcription, Genetic
D050957:HSP110 Heat-Shock Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins