1 Hebei Key Laboratory of Animal Diversity/College of Life Sciences, Langfang Normal University, Langfang 065000, China; 2 Key Laboratory of Animal (Poultry) Genetics Breeding and Reproduction, Ministry of Agriculture and Rural Affairs/Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Abstract：Heat shock transcription factor 1 (HSF1) is an important transcription factor that initiates heat shock protein gene expression. Further study on HSF1 will help to clarify the regulatory mechanism of HSF1 on heat stress response. In this study, reverse transcription-PCR (RT-PCR) was used to clone the full-length CDS of HSF1 from Chinese Holstein cow (Bos taurus), and bioinformatics was used to analyze the biological characteristics of the protein. The results showed that 3 splice variants were successfully cloned, named HSF1-AS1 (GenBank No. MW401766), HSF1-AS2 (GenBank No. XM_005215151.4), and HSF1-AS3 (GenBank No. MW401767). The splicing patterns of the 3 variants were exon skipping, intron retaining, and alternative 3' splice sites. The ORF analysis showed that the 3 splice variants of HSF1 encoded 311, 553 and 517 aa, respectively. The RNA-seq data of breast tissue in previous study was analyzed, and the results showed that the expression of HSF1-AS2 was the highest (P<0.01), and the expression of HSF1-AS1 was the lowest (P<0.01). The results of qRT-PCR showed that the expression levels of HSF1-AS1 and HSF1-AS2 were both up-regulated in heat stressed mammary epithelial cells. The binding site analysis of splicing factors showed that the SNP (18948 C>T) of exon 11 of HSF1 gene was just in the potential region of exonic splicing enhancer (ESE), which might be associated with the formation of HSF1-AS3. The phylogenetic analysis showed that bovine HSF1 amino acid sequence had high homology and close genetic distance with that of goats (Capra hircus). The present study provides a reference for further elucidating the regulation mechanism of HSF1 gene expression.
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