Complementation between mutants of CHO cells resistant to a variety of plant lectins

Stanley, Pamela; Siminovitch, Louis

doi:10.1007/bf01542968

Cited by 151 publications

(117 citation statements)

References 18 publications

Supporting

Mentioning

116

Contrasting

Order By: Relevance

“…For polyomaviruses, enzymatic or biochemical removal of SA from the host cell (1975); Stanley & Siminovitch (1977); this study 2* ( 2) 0?2 ND Stanley et al (1975); Stanley & Siminovitch (1977); this study 2* ( +) 21 ND Stanley et al (1975); Stanley & Siminovitch (1977); this study *Reduced amounts of SA on the cell surface. ND, Not determined.…”

Section: Discussionmentioning

confidence: 99%

“…Mock-or a4-transfected BALB/c 3T3 cells were grown in DMEM supplemented with 10 % FCS in the presence of 1?3 mg puromycin ml 21 , as described previously (Caruso et al, 2003). Lec-2 cells, defective in SA expression, and the parental cell line Pro-5 (expressing SA), both originating from a Chinese ovary hamster (CHO) cell line, were grown as monolayers, as described previously (Stanley et al, 1975;Stanley & Siminovitch, 1977). Pro-5 and Lec-2 cells were transfected with the empty or recombinant pRK5 vector carrying the entire cDNA of the murine a4 integrin subunit (Rietzler et al, 1998) and the plasmid pBabe-Puro containing the puromycin-resistance gene as a selection marker (Morgenstern & Land, 1990) using the Lipofectamine Plus reagent (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…The binding properties of mutant and wt VLPs were also compared in the CHO-derived cell lines Pro-5 and Lec-2. The Lec-2 cell line is a Pro-5-subline reported to express 10 % of the total amount of SA on the cell membrane with respect to the parental cell line (Stanley et al, 1975;Stanley & Siminovitch, 1977). First of all, we controlled by FACS the expression level of SA on both cell lines using biotinylated WGA, a lectin that interacts with N-linked a(2,3) and a(2,6) SA residues in a manner similar to that of VP1 (Stehle & Harrison, 1997).…”

Section: Cell Binding Of Mutant Vlps Is Totally Abolishedmentioning

confidence: 99%

See 2 more Smart Citations

Role of sialic acid-containing molecules and the α4β1 integrin receptor in the early steps of polyomavirus infection

Caruso

Cavaldesi

Gentile

et al. 2003

Journal of General Virology

View full text Add to dashboard Cite

Murine polyomavirus (MPyV) infection occurs through recognition of sialic acid (SA) residues present on the host cell membrane, but the nature of the molecules involved and the exact role of this interaction in virus cell entry still need to be clarified. In this work, mutations at residues R 77 or H 298 of the MPyV VP1 protein were shown to lead to a complete loss of virus infectivity, which, however, could be restored by lipofection of virus particles into the cytoplasm of the host cells. Using virus-like particles (VLPs), it was demonstrated that the non-infectivity of these mutants was due to impaired cell entry caused by total abrogation of SA-dependent cell binding. This indicates that SA residues are essential primary cell receptors for MPyV. As the a4b1 integrin has been identified recently as a cell receptor for MPyV, the relationship, if any, was investigated between SA-containing and a4b1 integrin receptors. The ability of mutants R 77 Q and H 298 Q and wt VLPs to bind to cells overexpressing the a4b1 integrin was studied in SA-positive (BALB/c 3T3 cells and Pro-5 cells) and SA-deficient (Pro5-derived Lec-2 cells) backgrounds. Overexpression of a4b1 integrin did not restore binding of mutant VLPs in any of these cell lines or, indeed, that of wt VLPs in a SA-deficient background. Moreover, evidence is provided that overexpression of the sialylated a4b1 integrin enhances wt VLP cell binding, suggesting that, in addition to its function at a post-attachment level, a4b1 integrin acts also as one of the SA-containing receptors for initial cell binding.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Cell Binding Of Mutant Vlps Is Totally Abolishedmentioning

confidence: 99%

See 1 more Smart Citation

Role of sialic acid-containing molecules and the α4β1 integrin receptor in the early steps of polyomavirus infection

Caruso

Cavaldesi

Gentile

et al. 2003

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…Materials-The Lec1-CHO cell line was developed by Dr. Pamela Stanley (53). COS7 and Lec1-CHO cells were from the American Type Culture Collection (Manassas, VA).…”

Section: Methodsmentioning

confidence: 99%

O-Glucose Trisaccharide Is Present at High but Variable Stoichiometry at Multiple Sites on Mouse Notch1

Rana¹,

Nita‐Lazar²,

Takeuchi

et al. 2011

Journal of Biological Chemistry

120

View full text Add to dashboard Cite

Notch activity is regulated by both O-fucosylation and O-glucosylation, and Notch receptors contain multiple predicted sites for both. Here we examine the occupancy of the predicted O-glucose sites on mouse Notch1 (mN1) using the consensus sequence C 1 XSXPC 2 . We show that all of the predicted sites are modified, although the efficiency of modifying O-glucose sites is site-and cell type-dependent. For instance, although most sites are modified at high stoichiometries, the site at EGF 27 is only partially glucosylated, and the occupancy of the site at EGF 4 varies with cell type. O-Glucose is also found at a novel, nontraditional consensus site at EGF 9. Based on this finding, we propose a revision of the consensus sequence for O-glucosylation to allow alanine N-terminal to cysteine 2: C 1 XSX(A/P)C 2 . We also show through biochemical and mass spectral analyses that serine is the only hydroxyamino acid that is modified with O-glucose on EGF repeats. The O-glucose at all sites is efficiently elongated to the trisaccharide Xyl-Xyl-Glc. To establish the functional importance of individual O-glucose sites in mN1, we used a cell-based signaling assay. Elimination of most individual sites shows little or no effect on mN1 activation, suggesting that the major effects of O-glucose are mediated by modification of multiple sites. Interestingly, elimination of the site in EGF 28, found in the Abruptex region of Notch, does significantly reduce activity. These results demonstrate that, like O-fucose, the O-glucose modifications of EGF repeats occur extensively on mN1, and they play important roles in Notch function.The Notch family of single-pass transmembrane receptors is essential for early metazoan development, activating the expression of many genes involved in cell differentiation and tissue morphogenesis (1-3). Defects in Notch signaling have been implicated in a number of human diseases, including several forms of cancer, vascular defects such as cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (4 -6), multiple sclerosis (6), and a number of developmental syndromes (4, 7-10). The canonical Notch signaling pathway is initiated by the interaction of Notch with its ligand on an apposed cell. Upon ligand binding, Notch undergoes presenilin-1-dependent proteolysis, releasing a soluble Notch intracellular domain, which enters the cell nucleus to interact directly with the transcription factors from the CSL family and regulate Notch target genes (2). There are four members of the Notch family in vertebrates (Notch1 to Notch4) interacting with several classes of Notch ligands: ligands of the DSL family (Delta-like 1, 3, and 4 and Jagged 1 and 2) (1) and newly characterized ligands without the DSL domain, such as DNER and MAGP-1 and -2 (11-13).The Notch proteins consist of a large extracellular domain (ECD), 5 a transmembrane region, and a large intracellular domain (1, 2, 14). The majority of the ECD consists of tandem epidermal growth factor-like (EGF) repeats (36 EGF repeats are ...

show abstract

“…Golgi membranes from uninfected Chinese hamster ovary (CHO) wild-type (Pro − 5 ) and VSV-infected mutant CHO cell lines Lec 1 (defective in GlcNAc transferase I; (67,68)), Lec 8 (defective in UDPgalactosyl translocase; (69)), and Lec 2 (defective in CMP-sialic acid translocase; (70)) were prepared as previously described (23). CHO cytosol was prepared as described by Block et al (71).…”

Section: Cell Culture and Biological Materialsmentioning

confidence: 99%

Coatomer Vesicles are not Required for Inhibition of Golgi Transport by G‐Protein Activators

Happe

Cairns

Roth

et al. 2000

Traffic

View full text Add to dashboard Cite

The G-protein activators guanosine 5%-O-(3-thiodiphosphate) (GTPgS) and aluminum fluoride (AlF) are thought to inhibit transport between Golgi cisternae by causing the accumulation of nonfunctional coatomer-coated transport vesicles on the Golgi. Although GTPgS and AlF inhibit transport in cell-free intra-Golgi transport systems, blocking coatomer vesicle formation does not. We therefore determined whether inhibition of in vitro Golgi transport by these agents requires coatomer vesicle formation. Depletion of coatomer was found to completely block coated vesicle formation on Golgi cisternae without affecting inhibition of in vitro transport by either GTPgS or AlF. Depletion of ADP-ribosylation factor (ARF) prevented inhibition of transport by GTPgS, but not by AlF, suggesting that the AlF-sensitive component in transport may not be a GTP-binding protein. Surprisingly, depletion of cytosolic ARF did not prevent the GTPgS-induced formation of Golgi-coated vesicles, whereas ARF was required for AlF-induced vesicle formation. Although ARF or coatomer depletion caused an increase in the fenestration of cisternae, no other utrastructural changes were observed that might explain the inhibition of transport by GTPgS or AlF. These findings suggest that ARF-GTPgS and AlF act by distinct and coatomer-independent mechanisms to inhibit membrane fusion in cell-free intra-Golgi transport.

show abstract

Complementation between mutants of CHO cells resistant to a variety of plant lectins

Cited by 151 publications

References 18 publications

Role of sialic acid-containing molecules and the α4β1 integrin receptor in the early steps of polyomavirus infection

Role of sialic acid-containing molecules and the α4β1 integrin receptor in the early steps of polyomavirus infection

O-Glucose Trisaccharide Is Present at High but Variable Stoichiometry at Multiple Sites on Mouse Notch1

Coatomer Vesicles are not Required for Inhibition of Golgi Transport by G‐Protein Activators

Contact Info

Product

Resources

About