Coding sequence and expression of the homeobox gene Hox 1.3

Development ◽  
1988 ◽  
Vol 102 (2) ◽  
pp. 349-359 ◽  
Author(s):  
M. Fibi ◽  
B. Zink ◽  
M. Kessel ◽  
A.M. Colberg-Poley ◽  
S. Labeit ◽  
...  
Keyword(s):  
Homeobox Gene ◽  
Cell System ◽  
T Antigen ◽  
Open Reading Frame ◽  
Homeodomain Protein ◽  
Chromosome 11 ◽  
3T3 Cells ◽  
Exon 2 ◽  
Reading Frame ◽  
3T3 Fibroblasts

We have characterized Hox 1.3 (previously described as m2), a murine homeobox-containing gene, which is a member of the Hox 1 cluster located on chromosome 6. A cloned cDNA was isolated from an Okayama-Berg library generated from the chemically transformed cell line MB66 MCA ACL6. The protein sequence of 270 amino acids was deduced from the nucleotide sequence of an open reading frame containing the homeobox. The open reading frame is interrupted at the genomic level by a 960 bp intron and is organized in two exons. The Hox 1.3 protein was found to contain extensive sequence homology with the murine homeodomain protein Hox 2.1, which is encoded on chromosome 11. There are two homology with the regions in the first exon, i.e. a hexapeptide conserved in many homeobox-containing genes and the N-terminal domain, which was found to be homologous only to Hox 2.1. Furthermore, in exon 2 the homologies of the homeodomain regions are extended up to the carboxy terminus of Hox 1.3 and Hox 2.1. During prenatal murine development, maximal expression of Hox 1.3 is observed in 12-day embryonic tissue. The two transcripts carrying the Hox 1.3 homeobox are 1.9 kb and about 4 kb in length. An abundant Hox 1.3-specific 1.9 kb RNA is also found in F9 cells which were induced for parietal endoderm differentiation, whereas F9 teratocarcinoma stem cells do not stably express this specific RNA. Induction of the transcript occurs immediately after retinoic acid/cAMP treatment and the RNA level remains high for 5 days. Thus, the kinetics are different from the previously described homeobox transcripts Hox 1.1 and Hox 3.1. Interestingly, by analogy to the F9 cell system a negative correlation between transformation and Hox 1.3 expression is observed in 3T3 fibroblasts also. Untransformed 3T3 cells carry abundant 1.9 kb Hox 1.3 RNA, whereas the methylcholanthrene-transformed MB66 and LTK- cells or 3T3 cells transformed by the oncogenes src, fos or SV40 T antigen express only low levels.

Oncogenic Transformation of NIH 3T3 Fibroblasts by BCR-ABL Oncoprotein Requires the IL-3 Receptor.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3370-3370
Author(s):  
Wenjing Tao ◽  
Hui Lin ◽  
Tong Sun ◽  
Ajoy K. Samanta ◽  
Ralph B. Arlinghaus
Keyword(s):  
Kinase Inhibitor ◽  
Philadelphia Chromosome ◽  
Cell System ◽  
Receptor Expression ◽  
Oncogenic Transformation ◽  
3T3 Cells ◽  
Lymphoid Leukemia ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Abstract Bcr-Abl is a leukemia-inducing protein, which causes oncogenic transformation of myeloid progenitors in Philadelphia chromosome (Ph)-positive chronic myeloid leukemia (CML) and lymphoid progenitors in Ph+ acute lymphoid leukemia (ALL). Oncogenic transformation of hematopoietic cells by the Bcr-Abl oncoprotein directly involves the activation Jak2 tyrosine kinase and the Stat5 transcription factor. Both proteins are normally linked to the IL-3/GM-CSF receptors for growth and survival. Since fibroblastic cells are not targets of BCR-ABL induced oncogenesis, we determined whether forced expression of the IL-3 receptor would allow oncogenic transformation of NIH 3T3 fibroblasts known to be resistant to transformation by BCR-ABL. NIH 3T3 cells transduced with the human IL-3 receptor a and b chains were highly susceptible to oncogenic transformation by expression of BCR-ABL. Forced expression of both receptor chains but not either one alone allowed efficient foci formation of NIH 3T3 cells expressing BCR-ABL, and these cells formed colonies in soft agar whereas BCR-ABL positive NIH 3T3 cells lacking IL-3 receptor expression did not. The Bcr-Abl kinase inhibitor imatinib mesylate (1 mM) and the Jak kinase inhibitor AG490 (10 mM) strongly inhibited agar colony formation. A small molecule inhibitor of Jak2 kinase, 1,2,3,4,5,6-hexabromocyclohexane reported to be specific for Jak2 (Sandberg et al. J. Med. Chem, 2005)-significantly reduced the phosphorylation of Gab2 at the YxxM motif, which is needed for activation of the PI-3 kinase and Akt, two proteins that are part of the Bcr-Abl/Jak2 Network (Samanta et al. Cancer Res, 2006). These findings indicate that Bcr-Abl oncoprotein requires the IL-3 receptor/Jak2/Stat5 pathways for oncogenic transformation of NIH 3T3 fibroblasts, and may explain partially why Bcr-Abl oncogenesis is restricted to hematopoietic malignancies. Furthermore, this cell system in fibroblastic and other cell lineages will provide a model to probe the detailed steps that require IL-3 receptor and Jak2 for Bcr-Abl induced leukemia.

Influence of amino acids encoded in the 3′ open reading frame of the SV40 early region on transformation and antigenicity of large T antigen

Virology ◽  
1986 ◽  
Vol 150 (2) ◽  
pp. 361-372 ◽  
Author(s):  
M.J. Tevethia ◽  
R.W. Anderson ◽  
S.S. Tevethia ◽  
D. Simmons ◽  
J. Feunteun ◽  
...  
Keyword(s):  
Amino Acids ◽  
T Antigen ◽  
Open Reading Frame ◽  
Large T Antigen ◽  
Reading Frame ◽  
Early Region ◽  
Sv40 Early Region

scholarly journals 5′ Heterogeneity in epithelial sodium channel α-subunit mRNA leads to distinct NH2-terminal variant proteins

1998 ◽  
Vol 274 (5) ◽  
pp. C1312-C1323 ◽  
Author(s):  
Christie P. Thomas ◽  
Scott Auerbach ◽  
John B. Stokes ◽  
Kenneth A. Volk
Keyword(s):  
Sodium Channel ◽  
Xenopus Oocytes ◽  
Epithelial Sodium Channel ◽  
Open Reading Frame ◽  
Splice Sites ◽  
Α Subunit ◽  
Exon 2 ◽  
Reading Frame ◽  
Subunit Mrna ◽  
Epithelial Sodium Channel Enac

The amiloride-sensitive epithelial sodium channel (ENaC) is composed of three subunits: α, β, and γ. The human α-ENaC subunit is expressed as at least two transcripts (N. Voilley, E. Lingueglia, G. Champigny, M. G. Mattei, R. Waldmann, M. Lazdunski, and P. Barbry. Proc. Natl. Acad. Sci. USA91: 247–251, 1994). To determine the origin of these transcripts, we characterized the 5′ end of the α-ENaC gene. Four transcripts that differ at their first exon were identified. Exon 1A splices to exon 2 to form the 5′ end of α-ENaC1, whereas exon 1B arises separately and continues into exon 2 to form α-ENaC2. Other variant mRNAs, α-ENaC3 and α-ENaC4, are formed by activating 5′ splice sites within exon 1B. Although α-ENaC3 and -4 did not change the open reading frame for α-ENaC, α-ENaC2 contains upstream ATGs that add 59 amino acids to the previous (α-ENaC1) protein. To address the significance of these isoforms, both proteins were expressed in Xenopus oocytes. The cRNA for each α-ENaC transcript when combined with β- and γ-ENaC cRNA reconstituted a low-conductance ion channel with amiloride-sensitive currents of similar characteristics. We have thus identified variant α-ENaC mRNAs that lead to functional ENaC peptides.

scholarly journals Human Polyomavirus-Encoded Circular RNAs

2020 ◽  
Author(s):  
Rong Yang ◽  
Eunice E. Lee ◽  
Jiwoong Kim ◽  
Joon H. Choi ◽  
Yating Chen ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Cultured Cells ◽  
Reporter Genes ◽  
T Antigen ◽  
Open Reading Frame ◽  
Human Polyomavirus ◽  
Circular Rnas ◽  
Large T Antigen ◽  
Reading Frame ◽  
Human Polyomaviruses

ABSTRACTCircular RNAs (circRNAs) are a conserved class of RNAs with diverse functions. A subset of circRNAs are translated into peptides. Here we describe circular RNAs encoded by human polyomaviruses (HPyVs), including circular forms of RNAs encoding variants of the previously described alternative large T antigen open reading frame (ALTO) gene. Circular ALTO RNAs (circALTOs) can be detected in virus positive Merkel cell carcinoma (VP-MCC) cell lines and tumor samples. CircALTOs are stable, predominantly located in the cytoplasm, and N6-methyladenosine (m6A) modified. MCPyV circALTOs produce ALTO protein in cultured cells. MCPyV ALTO promotes the transcription of co-transfected reporter genes. MCPyV circALTOs are enriched in exosomes derived from VP-MCC lines and circALTO-transfected 293T cells, and purified exosomes can mediate ALTO expression and transcriptional activation. The related trichodysplasia spinulosa polyomavirus (TSPyV) also expresses a circALTO that can be detected in infected tissues and produces ALTO protein in cultured cells. Thus, human polyomavirus circRNAs are expressed in human tumors and tissues, encode for proteins, and may contribute to the infectious and tumorigenic properties of these viruses.

scholarly journals Adenovirus Type 5 E4 Open Reading Frame 4 Protein Induces Apoptosis in Transformed Cells

1998 ◽  
Vol 72 (4) ◽  
pp. 2975-2982 ◽  
Author(s):  
Ronit Shtrichman ◽  
Tamar Kleinberger
Keyword(s):  
Adenovirus Type ◽  
Open Reading Frame ◽  
Wild Type ◽  
3T3 Cells ◽  
Reading Frame ◽  
293 Cells ◽  
Lung Carcinoma Cell Line ◽  
Phosphatase 2A ◽  
Nih 3T3 ◽  
Adenovirus Type 5

ABSTRACT Adenovirus type 5 E4 open reading frame 4 (E4orf4) protein has been previously shown to counteract transactivation of the junBand c-fos genes by cyclic AMP plus E1A protein and to interact with protein phosphatase 2A (PP2A). Here, we show that the wild-type E4orf4 protein induces apoptosis in the E1A-expressing 293 cells, in NIH 3T3 cells transformed with v-Ras, and in the lung carcinoma cell line H1299. The induction of apoptosis is not accompanied by enhanced levels of p53 in 293 cells and occurs in the absence of p53 in H1299 cells, indicating involvement of a p53-independent pathway. A mutant E4orf4 protein that had lost the ability to induce apoptosis also lost its ability to bind PP2A. We suggest that E4orf4 antagonizes continuous signals to proliferate, like those given by E1A or v-Ras, and that the conflicting signals lead to the induction of cell death.

Abstract 022: A Short Open Reading Frame in the Angiotensin Type 1a Receptor 5' Leader Sequence Increases the Rate of Receptor Internalization

Hypertension ◽  
2015 ◽  
Vol 66 (suppl_1) ◽  
Author(s):  
Parnika Kadam ◽  
Susette Mueller ◽  
Hong Ji ◽  
Kathryn Sandberg
Keyword(s):  
Laser Scanning ◽  
Leader Sequence ◽  
Open Reading Frame ◽  
Receptor Internalization ◽  
Start Codon ◽  
Ang Ii ◽  
Exon 2 ◽  
Reading Frame ◽  
Short Open Reading Frame ◽  
Angiotensin Type

Background: We recently found that a seven amino acid peptide (PEP7) encoded within a short open reading frame (sORF) in exon 2 of the 5’ leader sequence (5'LS) of the angiotensin type 1a receptor (AT1aR) mRNA inhibits AT1aR-mediated activation of extracellular signal-regulated protein kinases 1 and 2 (Erk1/2) without having any effect on the AT1aR-inositol trisphosphate protein kinase C pathway. To investigate the mechanism by which PEP7 selectively inhibits the AT1aR-Erk1/2 signaling cascade, the start codon of the sORF was mutated at adenine -108 (A-108 to T-108) to create E1,2(-108T),3-AT1aR followed by cloning the E1,2,3-AT1aR and the mutant E1,2(-108T),3-AT1aR into the pEGFP-N2 plasmid. Methods: Human embryonic kidney 293 (HEK-293) cells were transfected with E1,2,3-AT1aR (intact sORF construct) or E1,2(-108T),3-AT1aR (disrupted sORF construct) by lipofectamine. Forty eight to seventy two hours later, live cell time lapse images were collected before and after treatment with Ang II (100 nM) using a TE300 Spinning disk laser scanning confocal microscope. Image analysis was performed using Velocity software. Results: Within 5 min of Ang II stimulation, punctae formed and moved throughout the cell membrane in cells transfected with both EGFP tagged receptors; however, even though the punctae represent the localized accumulation of identical AT1aR proteins, there were distinct differences in the intensity and time course of punctae. The rate of vesicle formation after Ang II treatment was markedly decreased by disrupting the PEP7 sORF [t1/2 (s): E1,2,3-AT1aR, 203s (N=21) vs E1,2(-108T),3-AT1aR, 328s (N=14); p<0.0001]. Conclusion: The PEP7 sORF facilitates Ang II-induced AT1R internalization. These findings suggest that we have uncovered a new mechanism governing agonist-induced AT1aR cellular trafficking that could have implications not only for regulation of AT1aR signaling cascades but also for other trafficking proteins that contain an upstream sORF within their 5'LS.

scholarly journals Inhibition of Human Cytomegalovirus DNA Maturation by a Benzimidazole Ribonucleoside Is Mediated through the UL89 Gene Product

1998 ◽  
Vol 72 (1) ◽  
pp. 717-725 ◽  
Author(s):  
Mark R. Underwood ◽  
Robert J. Harvey ◽  
Sylvia C. Stanat ◽  
Mary Lou Hemphill ◽  
Teresa Miller ◽  
...  
Keyword(s):  
Gene Product ◽  
Human Cytomegalovirus ◽  
Bacteriophage T4 ◽  
Serial Passage ◽  
Open Reading Frames ◽  
Open Reading Frame ◽  
Viral Dna ◽  
Exon 2 ◽  
Reading Frame ◽  
Dependent Endonuclease

ABSTRACT 2-Bromo-5,6-dichloro-1-β-d-ribofuranosyl benzimidazole (BDCRB) is a member of a new class of benzimidazole ribonucleosides which inhibit human cytomegalovirus (HCMV) late in the replication cycle without inhibiting viral DNA synthesis. We show here that polygenomic concatemeric HCMV DNA does not mature to unit genome length in the presence of BDCRB. To discover the locus of action, virus resistant to BDCRB was selected by serial passage in the presence of the compound. Genetic mapping experiments with BDCRB-resistant virus demonstrated that the resistance phenotype mapped to one amino acid (Asp344Glu; low resistance) or two amino acids (Asp344Glu and Ala355Thr; high resistance) within the product of exon 2 of the HCMV UL89 open reading frame. The HCMV UL89 open reading frame and its homologs are among the most conserved open reading frames in the herpesviruses, and their products have sequence similarities to a known ATP-dependent endonuclease from the double-stranded DNA bacteriophage T4. These findings strongly suggest that BDCRB prevents viral DNA maturation by interacting with a UL89 gene product and that the UL89 open reading frame may encode an endonucleolytic subunit of the putative HCMV terminase. Further, since mammalian cell DNA replication does not involve a DNA maturation step, compounds which inhibit viral DNA maturation should be selective and safe.

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