Cloning, Expression and Functional Role of XRPFI α and β Subunits in Xenopus laevis Oocyte

Gorgoni, Barbara; Fiorentino, L.; Marchioni, Marcella; Carnevali, Francesca

doi:10.1006/bbrc.1995.2575

Cited by 5 publications

(2 citation statements)

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“…For the oocyte swelling assays, the pT7Ts‐derived vector containing the T7 RNA polymerase promoter and carrying the 5′ and 3′ translated regions of the Xenopus laevis β‐globin gene (Gorgoni et al ., 1995) was modified to add new cloning sites. Briefly, overlapping oligos with the recognition sites for Not I, Xho I, and Sac II (Table S1) were annealed and cloned into the vector previously linearised using the Bgl II and Spe I enzymes.…”

Section: Methodsmentioning

confidence: 99%

Plasma membrane aquaporins interact with the endoplasmic reticulum resident VAP27 proteins at ER–PM contact sites and endocytic structures

et al. 2020

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Plasma membrane (PM) intrinsic proteins (PIPs) are aquaporins facilitating the diffusion of water and small solutes. The functional importance of the PM organisation of PIPs in the interaction with other cellular structures is not completely understood. We performed a pull-down assay using maize (Zea mays) suspension cells expressing YFP-ZmPIP2;5 and validated the protein interactions by yeast split-ubiquitin and bimolecular fluorescence complementation assays. We expressed interacting proteins tagged with fluorescent proteins in Nicotiana benthamiana leaves and performed water transport assays in oocytes. Finally, a phylogenetic analysis was conducted. The PM-located ZmPIP2;5 physically interacts with the endoplasmic reticulum (ER) resident ZmVAP27-1. This interaction requires the ZmVAP27-1 cytoplasmic major sperm domain. ZmPIP2;5 and ZmVAP27-1 localise in close vicinity in ER-PM contact sites (EPCSs) and endocytic structures upon exposure to salt stress conditions. This interaction enhances PM water permeability in oocytes. Similarly, the Arabidopsis ZmVAP27-1 paralogue, AtVAP27-1, interacts with the AtPIP2;7 aquaporin. Together, these data indicate that the PIP2-VAP27 interaction in EPCSs is evolutionarily conserved, and suggest that VAP27 might stabilise the aquaporins and guide their endocytosis in response to salt stress.

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Section: Methodsmentioning

confidence: 99%

Plasma membrane aquaporins interact with the endoplasmic reticulum resident VAP27 proteins at ER–PM contact sites and endocytic structures

et al. 2020

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“…For preparation of in-vitro made RNA specific for FIII/YY1, the translated region (nucleotides ϩ49 to ϩ1167) of the FIII/ YY1 cloned cDNA was amplified by PCR, blunt-ended and cloned into the unique EcoRV site of the T7-RNA-polymerase directed T7TS vector, a derivative of pGEM4Z (Promega) with 5′ and 3′ untranslated regions of Xenopus β-globin mRNA plus 30 poly(A) and 30 poly(C) residues [19]. HindIII-linearized T7TS-FIII was transcribed in vitro with the T7 RNA polymerase kit (Boehringer) with trace amounts of [ 32 P]UTP (Amersham).…”

Section: Methodsmentioning

confidence: 99%

The binding sites for Xenopus laevis FIII/YY1 in the first exon of L1 and L14 ribosomal protein genes are dispensable for promoter expression

Rinaldis

Pisaneschi²,

Camacho-Vanegas

et al. 1998

European Journal of Biochemistry

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The genes coding for the ribosomal proteins (rp genes) L14 and L1 in the toad Xenopus laevis are contacted in the first exon by the frog protein, FIII/YY1, homolog of the human zinc-finger protein YY1, acting as repressor, activator and initiator of transcription. To investigate the functional significance of FIII/YY1 in the context of the two rp genes, the L14 region at nucleotide positions Ϫ105 to ϩ44, including all of the first exon was linked to the chloramphenicol acetyltransferase (CAT) reporter gene; constructs with wild-type and mutated sites for FIII/YY1 were injected into nuclei of stage VϪVI oocytes and analyzed for CAT activity. The same procedure was followed for constructs made with L1 sequences at nucleotide positions Ϫ17 to ϩ1567. Mutations in the sites for FIII/YY1 did not change reporter activity, nor did overexpression of FIII/YY1 in the oocytes prior to injection with L1 and L14 constructs. Since oocytes are non-dividing cells, transfections were made of Xenopus kidney cells in culture with the same constructs and the results obtained in oocytes confirmed.Keywords : Xenopus laevis; ribosomal protein L1; ribosomal protein L14; YY1; FIII/YY1.The genes coding for vertebrate ribosomal proteins share some structural features: in general, they lack a canonical TATA box and the initiation of transcription is inside a run of consecutive pyrimidine residues. Common to a number of ribosomal protein genes (rp genes) is also the presence of a consensus binding site for the multifunctional factor YY1 in a position internal to the gene, often close to the ATG sequence for the initiator methionine residue. YY1 is a zinc-finger DNA-binding protein expressed in most, if not all, cell types. The human [1] and murine [2Ϫ4] cDNAs have been cloned during studies of transcriptional regulation. YY1 represses transcription in a variety of cellular and viral promoters [5,6], but acts as an activator when located near the site for initiation of transcription [7Ϫ9].In Xenopus laevis it was confirmed [10], by methylationinterference analysis, that the consensus sites in the first exon of L1 and L14 ribosomal protein genes are indeed contacted by the frog homolog, FIII/YY1, present in oocyte nuclei or overexpressed in oocytes by the cloned cDNA [10].Our previous studies of the untranscribed region of the X. laevis L14 rp gene promoter evidenced the importance of Sp1 and of XrpFI (the homolog of rat GABP A and β heterodimer [11]) for the expression of the gene in oocyte nuclei and in Hela cells [12,13]. When the upstream region with the Sp1 site atCorrespondence to E. Beccari, Centro di Studio per gli Acidi Nucleici CNR, Dipartimento di Genetica e Biologia Molecolare, Università di Roma La Sapienza, P.le A. Moro 5, I-00185 Roma, Italy Phone: ϩ39 6 49912337. Fax: ϩ39 6 49912500. E-mail: beccarie@axrma.uniroma1.it Abbreviations. CAT, chloramphenicol acetyltransferase; GST, glutathione S-transferase; RSV, Rous sarcoma virus ; CMV, cytomegalovirus; luc, luciferase ; rp, ribosomal protein.Note. The L1 and L14 Xenopus laevis ri...

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Isolation and characterization of chicken GA-binding protein

Toku

Quaye

Tanaka

2002

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expres

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Cloning, Expression and Functional Role of XRPFI α and β Subunits in Xenopus laevis Oocyte

Cited by 5 publications

References 0 publications

Plasma membrane aquaporins interact with the endoplasmic reticulum resident VAP27 proteins at ER–PM contact sites and endocytic structures

Plasma membrane aquaporins interact with the endoplasmic reticulum resident VAP27 proteins at ER–PM contact sites and endocytic structures

The binding sites for Xenopus laevis FIII/YY1 in the first exon of L1 and L14 ribosomal protein genes are dispensable for promoter expression

Isolation and characterization of chicken GA-binding protein

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