Pregnancy and interferon-τ upregulate gene expression of members of the 1-8 family in the bovine uterus

• Published in: Biology of reproduction 2001-11, Vol.65 (5), p.1471-1480
• Main Authors: PRU, James K, AUSTIN, Kathy J, HAAS, Arthur L, HANSEN, Thomas R
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; Cattle - genetics; DNA, Complementary - chemistry; Female; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; In Situ Hybridization; Interferon Type I - pharmacology; Mammalian female genital system; Membrane Proteins; Molecular Sequence Data; Morphology. Physiology; Pregnancy; Pregnancy Proteins - pharmacology; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - genetics; Uterus - metabolism; Vertebrates: reproduction
• ISSN: 0006-3363; 1529-7268
• DOI: 10.1043/0006-3363(2001)065(1471:PAIUGE)2.0.CO;2

The 1-8 family (1-8U, 1-8D, Leu-13/9-27) of interferon (IFN)-inducible genes encodes proteins that are components of multimeric complexes involved with transduction of antiproliferative and homotypic adhesion signals. Human 1-8 family members are highly similar and are regulated by type 1 and type 2 IFNs. Because the bovine uterus is bathed in conceptus-derived IFN tau during early pregnancy, it was hypothesized that members of the 1-8 family were upregulated in the bovine uterus during early pregnancy. Oligonucleotide primers were designed based on human and rat 1-8U and Leu-13 cDNAs and used in reverse transcription polymerase chain reactions to amplify bovine cDNAs from endometrial RNA. The bovine 1-8U cDNA was sequenced, found to be 84% identical to the human 1-8U, and used to screen a bovine endometrial cDNA library to isolate the full-length 1-8U and Leu-13 cDNAs. The inferred amino acid sequences of bovine 1-8U and Leu-13 were 72% and 73% identical to their respective human counterparts. Bovine 1-8U and Leu-13 retain an amino acid motif that is conserved in other 1-8 family members and in some ubiquitin-conjugating enzymes (E2s). This motif is critical for function of E2s in covalently linking ubiquitin to targeted proteins. Northern blotting revealed that bovine endometrial 1-8U and Leu-13 mRNAs were upregulated on Day 15 of pregnancy (P < 0.0001) and continued to accumulate through Day 18 of pregnancy (P < 0.05) when compared with endometrium from nonpregnant cows. The bovine 1-8U and Leu-13 mRNAs were also upregulated (P < 0.05) by IFN tau (25 nM) within 3 h, continued to accumulate through 12 h, and reached a plateau at 12-24 h in cultured bovine endometrial cells. In situ hybridization revealed that mRNAs encoding 1-8 family members were heavily localized to glandular epithelium but also were present to a lesser extent in the luminal epithelium and stroma. The temporal upregulation of 1-8U and Leu-13 mRNAs by pregnancy and IFN tau and tissue distribution of these mRNAs paralleled closely that of the ubiquitin homolog, IFN-stimulated gene product 17. These IFN-induced proteins probably work together to prepare the endometrium for adhesion of the developing conceptus.