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Identification of biotin‐ and holocarboxylase synthetase‐dependent microRNAs in human fibroblasts
Author(s) -
Wijeratne Subhashinee Samudra,
Malkaram Sridhar Achary,
Zempleni Janos,
Pabian Michael Dylan,
Granatowicz Andrew Daniel
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.647.3
Subject(s) - biotin , fibroblast , microbiology and biotechnology , mutant , chromatin , biology , gene expression , cell culture , microrna , chemistry , biochemistry , gene , genetics
MicroRNAs (miRs) regulate their target genes by binding to the 3′‐UTR. Previously we reported that the expression of miR‐539 depends on biotin, thereby regulating the expression of holocarboxylase synthetase (HCS) in human kidney cells. HCS appears to be part of a multiprotein gene repression complex in human chromatin. We hypothesized that both biotin and HCS play important roles in regulating miRs. IMR‐90 primary human fibroblasts were cultured in media containing deficient, normal, and high biotin concentrations; in addition, HCS mutant fibroblast were cultured in high biotin medium to mimic the scenario in HCS‐deficient patients receiving biotin supplements. miRs were sequenced using the SOLiD platform. Expression analyses were performed using edgeR, R software. The expression of miR‐378 and miR‐34a* was greater in biotin‐deficient and ‐supplemented IMR‐90 cells, respectively, compared with biotin‐normal controls (p < 0.01). The expression of miR‐2110 and miR‐150 was lower, in biotin‐deficient and supplemented cells, respectively, compared with controls (p < 0.01). The expression of 58 miRs increased or decreased in HCS‐mutant fibroblasts compared to IMR‐90 cells in biotin‐normal medium (p < 0.01). We conclude that HCS activity is of greater importance than biotin supply in regulating miR expression. (SSW and SAM contributed equally.) (UNL ARD Hatch Act, NIH DK063945 , DK077816 , and DK082476 )