Analysis of Recombinant Human ADP-Ribosylation Factors by Reversed-Phase High-Performance Liquid Chromatography and Electrospray Mass Spectrometry

Berger, Scott J.; Claude, Alejandro; Melançon, Paul

doi:10.1006/abio.1998.2821

Cited by 5 publications

(3 citation statements)

References 61 publications

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“…The new cleavage sites conferred by these tags were confirmed for some selectants by constructing plasmids that overproduce the tagging products (Table I) traditionally, from single unbroken mRNAs with stop codons. The three selectants that encode Cys residues produced reporters whose masses exceeded the expected values by 305 Da, suggesting a disulfide linkage to a glutathione moiety (Berger et al ., 1998). This interpretation was supported by the loss of the extra 305 Da for all Cys‐containing protein species after treatment with dithiothreitol (DTT).…”

Section: Resultsmentioning

confidence: 99%

Resuming translation on tmRNA: a unique mode of determining a reading frame

1999

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The bacterial ribosome switches from an mRNA lacking an in-frame stop codon and resumes translation on a specialized RNA known as tmRNA, SsrA or 10Sa RNA. We find that the ribosome can reach and use the extreme 3Ј terminal codon of the defective mRNA prior to switching. The first triplet to be translated in tmRNA (the resume codon) is determined at two levels: distant elements in tmRNA restrict resume codon choice to a narrow window and local upstream elements provide precision. Insights from a randomizationselection experiment secure the alignment of tmRNA sequences from diverse species. The triplet UA(A/G) (normally recognized as a stop codon by release factor-1) is strongly conserved two nucleotides upstream of the resume codon. The central adenosine of this triplet is essential for tmRNA activity. The reading frame of tmRNA is determined differently from all other known reading frames in that the first translated codon is not specified by a particular tRNA anticodon.

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Section: Resultsmentioning

confidence: 99%

Resuming translation on tmRNA: a unique mode of determining a reading frame

1999

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“…Additional vectors encoding untagged or hemagglutinin (HA)-tagged forms of Arf3 were constructed by inserting a PCR fragment into the XhoI and KpnI sites of the pcDNA4/TO vector (Invitrogen) modified by inversion of the multiple cloning site. The hArf3 encoding fragment was generated using pET21d-Arf3 (Berger, 1998) as template. The construction of HA-tagged Arf3 involved first insertion of annealed oligonucleotides encoding the HA epitope followed by a stop codon between the KpnI and HindIII sites of pcDNA4/TO(-).…”

Section: Methodsmentioning

confidence: 99%

Arf3 Is Activated Uniquely at thetrans-Golgi Network by Brefeldin A-inhibited Guanine Nucleotide Exchange Factors

Manolea

Chun

Chen

et al. 2010

MBoC

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“…Cultures of BL21(DE3) harboring plasmids encoding human ARF5 (ARF5:PET21d ϩ ; Berger et al, 1998) and yeast N -myristoyl transferase (pBB131; Duronio et al, 1990) containing 100 g/ml ampicillin and 20 g/ml kanamycin were induced by addition of IPTG (0.5 mM) at an OD 600 of 0.8. Bacteria were harvested by centrifugation after 4 h of growth at ambient temperature.…”

Section: Purification Of Recombinant Myristoylated Arf5mentioning

confidence: 99%

GBF1: A Novel Golgi-associated BFA-resistant Guanine Nucleotide Exchange Factor That Displays Specificity for ADP-ribosylation Factor 5

Claude¹

1999

The Journal of Cell Biology

111

145

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Abstract. Expression cloning from a cDNA library prepared from a mutant CHO cell line with Golgi-specific resistance to Brefeldin A (BFA) identified a novel 206-kD protein with a Sec7 domain termed GBF1 for Golgi BFA resistance factor 1. Overexpression of GBF1 allowed transfected cells to maintain normal Golgi morphology and grow in the presence of BFA. Golgienriched membrane fractions from such transfected cells displayed normal levels of ADP ribosylation factors (ARFs) activation and coat protein recruitment that were, however, BFA resistant. Hexahistidinetagged-GBF1 exhibited BFA-resistant guanine nucleotide exchange activity that appears specific towards ARF5 at physiological Mg 2 ϩ concentration. Characterization of cDNAs recovered from the mutant and wildtype parental lines established that transcripts in these cells had identical sequence and, therefore, that GBF1 was naturally BFA resistant. GBF1 was primarily cytosolic but a significant pool colocalized to a perinuclear structure with the ␤ -subunit of COPI. Immunogold labeling showed highest density of GBF1 over Golgi cisternae and significant labeling over pleiomorphic smooth vesiculotubular structures. The BFA-resistant nature of GBF1 suggests involvement in retrograde traffic.

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Analysis of Recombinant Human ADP-Ribosylation Factors by Reversed-Phase High-Performance Liquid Chromatography and Electrospray Mass Spectrometry

Cited by 5 publications

References 61 publications

Resuming translation on tmRNA: a unique mode of determining a reading frame

Resuming translation on tmRNA: a unique mode of determining a reading frame

Arf3 Is Activated Uniquely at thetrans-Golgi Network by Brefeldin A-inhibited Guanine Nucleotide Exchange Factors

GBF1: A Novel Golgi-associated BFA-resistant Guanine Nucleotide Exchange Factor That Displays Specificity for ADP-ribosylation Factor 5

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