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Developmentally important proteins that are crucial for fertilization and embryogenesis are synthesized through highly regulated translation of maternal mRNA. The Zygote arrest proteins, Zar1 and Zar2, are crucial for embryogenesis and have been implicated in binding mRNA and repressing mRNA translation. To investigate Zar1 and Zar2, the full-length proteins had been fused to glutathione-S-transferase (GST) or MS2 protein tags with minimal inter-domain linkers derived from multiple cloning sites; however, these fusion proteins expressed poorly and/or lacked robust function. Here, we tested the effect of inserting additional linkers between the fusion domains. Three linkers were tested, each 17 amino acids long with different physical and chemical properties: flexible hydrophilic, rigid extended or rigid helical. In the presence of any of the three linkers, GST-Zar1 and GST-Zar2 had fewer breakdown products. Moreover, in the presence of any of the linkers, MS2-Zar1 was expressed to higher levels, and in dual luciferase tethered assays, both MS2-Zar1 and MS2-Zar2 repressed luciferase translation to a greater extent. These data suggest that for Zar fusion proteins, increasing the length of linkers, regardless of their physical or chemical properties, improves stability, expression and bioactivity.
Amet,
Insertion of the designed helical linker led to increased expression of tf-based fusion proteins.
2009, Pubmed
Amet,
Insertion of the designed helical linker led to increased expression of tf-based fusion proteins.
2009,
Pubmed
Arai,
Design of the linkers which effectively separate domains of a bifunctional fusion protein.
2001,
Pubmed
Argos,
An investigation of oligopeptides linking domains in protein tertiary structures and possible candidates for general gene fusion.
1990,
Pubmed
Bergeron,
Self-renaturing enzymes: design of an enzyme-chaperone chimera as a new approach to enzyme stabilization.
2009,
Pubmed
Bhandari,
1H-NMR study of mobility and conformational constraints within the proline-rich N-terminal of the LC1 alkali light chain of skeletal myosin. Correlation with similar segments in other protein systems.
1986,
Pubmed
Charlesworth,
The temporal control of Wee1 mRNA translation during Xenopus oocyte maturation is regulated by cytoplasmic polyadenylation elements within the 3'-untranslated region.
2000,
Pubmed
,
Xenbase
Charlesworth,
Musashi regulates the temporal order of mRNA translation during Xenopus oocyte maturation.
2006,
Pubmed
,
Xenbase
Charlesworth,
Xenopus laevis zygote arrest 2 (zar2) encodes a zinc finger RNA-binding protein that binds to the translational control sequence in the maternal Wee1 mRNA and regulates translation.
2012,
Pubmed
,
Xenbase
Chen,
Fusion protein linkers: property, design and functionality.
2013,
Pubmed
Coller,
mRNA stabilization by poly(A) binding protein is independent of poly(A) and requires translation.
1998,
Pubmed
Dankert,
Pre- and postovulatory aging of murine oocytes affect the transcript level and poly(A) tail length of maternal effect genes.
2014,
Pubmed
Dumont,
Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals.
1972,
Pubmed
,
Xenbase
Farley,
Regulation of maternal mRNAs in early development.
2008,
Pubmed
George,
An analysis of protein domain linkers: their classification and role in protein folding.
2002,
Pubmed
Gray,
Multiple portions of poly(A)-binding protein stimulate translation in vivo.
2000,
Pubmed
,
Xenbase
Hanahan,
Plasmid transformation of Escherichia coli and other bacteria.
1991,
Pubmed
Howard,
The mitogen-activated protein kinase signaling pathway stimulates mos mRNA cytoplasmic polyadenylation during Xenopus oocyte maturation.
1999,
Pubmed
,
Xenbase
Hu,
Mouse ZAR1-like (XM_359149) colocalizes with mRNA processing components and its dominant-negative mutant caused two-cell-stage embryonic arrest.
2010,
Pubmed
Huston,
Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli.
1988,
Pubmed
Jo,
The beneficial effects of antifreeze proteins in the vitrification of immature mouse oocytes.
2012,
Pubmed
Keryer-Bibens,
Tethering of proteins to RNAs by bacteriophage proteins.
2008,
Pubmed
Kim,
Mammalian Staufen1 recruits Upf1 to specific mRNA 3'UTRs so as to elicit mRNA decay.
2005,
Pubmed
Lee,
Enhanced antibacterial activity of an attacin-coleoptericin hybrid protein fused with a helical linker.
2013,
Pubmed
Lu,
Bifunctional enhancement of a beta-glucanase-xylanase fusion enzyme by optimization of peptide linkers.
2008,
Pubmed
MacNicol,
pXen, a utility vector for the expression of GST-fusion proteins in Xenopus laevis oocytes and embryos.
1997,
Pubmed
,
Xenbase
Machaca,
Induction of maturation-promoting factor during Xenopus oocyte maturation uncouples Ca(2+) store depletion from store-operated Ca(2+) entry.
2002,
Pubmed
,
Xenbase
Maeda,
Expression of a bifunctional chimeric protein A-Vargula hilgendorfii luciferase in mammalian cells.
1996,
Pubmed
Maeda,
Engineering of functional chimeric protein G-Vargula luciferase.
1997,
Pubmed
McCormick,
Immunization with an interferon-gamma-gp120 fusion protein induces enhanced immune responses to human immunodeficiency virus gp120.
2001,
Pubmed
Minshall,
A conserved role of a DEAD box helicase in mRNA masking.
2001,
Pubmed
,
Xenbase
Minshall,
Translational control assessed using the tethered function assay in Xenopus oocytes.
2010,
Pubmed
,
Xenbase
Misra,
Cell cycle-dependent regulation of the bi-directional overlapping promoter of human BRCA2/ZAR2 genes in breast cancer cells.
2010,
Pubmed
Nakajima,
Expression cloning of Xenopus zygote arrest 2 (Xzar2) as a novel epidermalization-promoting factor in early embryos of Xenopus laevis.
2009,
Pubmed
,
Xenbase
Nath,
Impact of gonadotropin supplementation on the expression of germ cell marker genes (MATER, ZAR1, GDF9, and BMP15) during in vitro maturation of buffalo (Bubalus bubalis) oocyte.
2013,
Pubmed
Pennetier,
Spatio-temporal expression of the germ cell marker genes MATER, ZAR1, GDF9, BMP15,andVASA in adult bovine tissues, oocytes, and preimplantation embryos.
2004,
Pubmed
Romar,
Expression of maternal transcripts during bovine oocyte in vitro maturation is affected by donor age.
2011,
Pubmed
SenGupta,
A three-hybrid system to detect RNA-protein interactions in vivo.
1996,
Pubmed
Shinojima,
Identification and analysis of an early diagnostic marker for malignant melanoma: ZAR1 intra-genic differential methylation.
2010,
Pubmed
Smith,
Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.
1988,
Pubmed
Summers,
Altered theca and cumulus oocyte complex gene expression, follicular arrest and reduced fertility in cows with dominant follicle follicular fluid androgen excess.
2014,
Pubmed
Wu,
Zygote arrest 1 (Zar1) is a novel maternal-effect gene critical for the oocyte-to-embryo transition.
2003,
Pubmed
Yamamoto,
Zar1 represses translation in Xenopus oocytes and binds to the TCS in maternal mRNAs with different characteristics than Zar2.
2013,
Pubmed
,
Xenbase
Zhao,
Increasing the homogeneity, stability and activity of human serum albumin and interferon-alpha2b fusion protein by linker engineering.
2008,
Pubmed
