title: Discovery of a novel oocyte-specific Krüppel-associated box domain-containing zinc finger protein required for early embryogenesis in cattle.
abstract: Zinc finger (ZNF) transcription factors interact with DNA through zinc finger motifs and play important roles in a variety of cellular functions including cell growth, proliferation, development, apoptosis, and intracellular signal transduction. One-third of ZNF proteins in metazoans contain a highly conserved N-terminal motif known as the Krüppel-associated box (KRAB) domain, which acts as a potent, DNA-binding dependent transcriptional repression module. Analysis of RNA-Seq data generated from a bovine oocyte cDNA library identified a novel transcript, which encodes a KRAB-containing ZNF transcription factor (named ZNFO). Characterization of ZNFO mRNA expression revealed that it is exclusively expressed in bovine oocytes and early embryos. A GFP reporter assay demonstrated that ZNFO protein localizes specifically to the nucleus, supporting its role in transcriptional regulation. To test the role of ZNFO in early embryonic development, zygotes were generated by in vitro maturation and fertilization of oocytes, and injected with small interfering RNA (siRNA) designed to knockdown ZNFO. Cleavage rates were not affected by ZNFO siRNA injection. However, embryonic development to 8- to 16-cell stage and blastocyst stage was significantly reduced relative to the uninjected and negative control siRNA-injected embryos. Further, interaction of ZNFO with the highly conserved co-factor, KRAB-associated protein-1 (KAP1), was demonstrated, and evidence supporting transcriptional repression by ZNFO was demonstrated using a GAL4-luciferase reporter system. Results of described studies demonstrate that ZNFO is a maternally-derived oocyte-specific nuclear factor required for early embryonic development in cattle, presumably functioning by repressing transcription.
authors.affiliation: Laboratory of Animal Biotechnology and Genomics, Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV 26506, United States.
authors.forename: Jacqelyn M
authors.initials: JM
authors.lastname: Hand
authors.affiliation: Laboratory of Mammalian Reproductive Biology and Genomics, Departments of Animal Science and Physiology, Michigan State University, East Lansing, MI 48824, United States.
authors.forename: Kun
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Laboratory of Animal Biotechnology and Genomics, Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV 26506, United States.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Laboratory of Animal Biotechnology and Genomics, Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV 26506, United States.
authors.forename: Prasanthi P
authors.initials: PP
authors.lastname: Koganti
authors.affiliation: Laboratory of Animal Biotechnology and Genomics, Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV 26506, United States.
authors.forename: Kristen
authors.initials: K
authors.lastname: Mastrantoni
authors.affiliation: Laboratory of Mammalian Reproductive Biology and Genomics, Departments of Animal Science and Physiology, Michigan State University, East Lansing, MI 48824, United States.
authors.forename: Sandeep K
authors.initials: SK
authors.lastname: Rajput
authors.affiliation: Laboratory of Mammalian Reproductive Biology and Genomics, Departments of Animal Science and Physiology, Michigan State University, East Lansing, MI 48824, United States; Department of Theriogenology, Faculty of Veterinary Medicine, Cairo University, Giza 12211, Egypt.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Ashry
authors.affiliation: Laboratory of Mammalian Reproductive Biology and Genomics, Departments of Animal Science and Physiology, Michigan State University, East Lansing, MI 48824, United States.
authors.forename: George W
authors.initials: GW
authors.lastname: Smith
authors.affiliation: Laboratory of Animal Biotechnology and Genomics, Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV 26506, United States. Electronic address: jianbo.yao@mail.wvu.edu.
authors.forename: Jianbo
authors.initials: J
authors.lastname: Yao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D047108:Embryonic Development; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D057809:HEK293 Cells; D006801:Humans; D009865:Oocytes / Q000502:physiology*; D021381:Protein Transport; D011817:Rabbits; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D012097:Repressor Proteins
keywords: Cattle; Embryogenesis; KRAB domain; Oocyte; Zinc finger