Polymorphisms in the bovine
HSP90AB1 gene are associated with heat tolerance in Thai indigenous cattle.
This notion could be further supported by mutations found in POLR1B (Table 1), which positively regulates gene transcription through epigenetic mechanism(s) [20] and
HSP90AB1 (Table 1), which modulates activity of several epigenetic regulators [21].
Hsp90ab1 was used as a housekeeping gene for data normalization in the Mouse Lipoprotein Signaling and Cholesterol Metabolism PCR Arrays.
Analysis of protein-protein interaction: STRING prediction indicated that UTY interacts with heat shock protein 90 kDa alpha, class A member 1 (HSP90AA1); heat shock protein 90 kDa alpha, class B member 1 (
HSP90AB1); ubiquitin specific peptidase 9, Y-linked (USP9Y); lysine (K)-specific demethylase 5D (KDM5D); histone deacetylase 2 (HDAC2); histone deacetylase 1 (HDAC1); histone deacetylase 3 (HDAC3); SET domain containing 2 (SETD2); DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, Y-linked (DDX3Y) and sex determining region Y (SRY) (Figure 4).
Results were normalized using at least two reference genes (GAPDH, HPRT1, ACTB, or
HSP90AB1) and were calculated using the [2.sup.-[DELTA][DELTA]C]T method [24].
Target gene amplifications were normalized using 3 validated housekeeping genes, ATP synthase O subunit (ATP5O), Glucuronidase Beta (GUSB), and Heat Shock Protein 90 Alpha Family Class B Member (
HSP90AB1) [15].
As with the low template concentration, PP1H (SD = 0.77), NONO (SD = 0.84), and GUSB (SD = 0.84) were the most stable of all genes tested, followed by RPLP0 and
HSP90AB1 (both with SD=1.02).
The expression of our candidate genes could be modulated mainly by 3 molecules: 5-hydroxy tryptamine (for HTR1B and SLC6A4), PRL (HTR2A and HTR2C), and
HSP90AB1 (for SL6CA4).
Following cell incubation with ox-VLDL, the
HSP90AB1, GLA, GCH1, and ARG2 genes are upregulated, whereas the DDAH2 and the GCHFR genes are downregulated (see Figure 1(c)).
HSP90AB1 was upregulated both in total cell extract analysis (fold change 2.3, p < 0.05, Student's f-test) (Figure S5 A) and in secretome analysis (fold change 4.8, p < 0.05, Student's f-test) (Figure S5 b) in C33A compared to HCK1T.
Using the same method, seven highly ranked genes (BAP1, GRB14,
HSP90AB1, ITGA5, NCKAP5L, SP1, and TOMM5) within [+ or -] 1 Mb of obesity SNPs were identified (Table 2).
Our data also exhibited that the stress-inducible protein-1 (STPI1) regulated
HSP90AB1 and HSPD1.