skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1378281
Resource Type:
Journal Article: Published Article
Journal Name:
Cell Reports
Additional Journal Information:
Journal Volume: 16; Journal Issue: 8; Related Information: CHORUS Timestamp: 2017-09-02 09:31:12; Journal ID: ISSN 2211-1247
Publisher:
Elsevier
Country of Publication:
Netherlands
Language:
English

Citation Formats

Marshall, Stephen S., Niesen, Michiel J. M., Müller, Axel, Tiemann, Katrin, Saladi, Shyam M., Galimidi, Rachel P., Zhang, Bin, Clemons, Jr., William M., and Miller, III, Thomas F. A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency. Netherlands: N. p., 2016. Web. doi:10.1016/j.celrep.2016.07.042.
Marshall, Stephen S., Niesen, Michiel J. M., Müller, Axel, Tiemann, Katrin, Saladi, Shyam M., Galimidi, Rachel P., Zhang, Bin, Clemons, Jr., William M., & Miller, III, Thomas F. A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency. Netherlands. doi:10.1016/j.celrep.2016.07.042.
Marshall, Stephen S., Niesen, Michiel J. M., Müller, Axel, Tiemann, Katrin, Saladi, Shyam M., Galimidi, Rachel P., Zhang, Bin, Clemons, Jr., William M., and Miller, III, Thomas F. 2016. "A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency". Netherlands. doi:10.1016/j.celrep.2016.07.042.
@article{osti_1378281,
title = {A Link between Integral Membrane Protein Expression and Simulated Integration Efficiency},
author = {Marshall, Stephen S. and Niesen, Michiel J. M. and Müller, Axel and Tiemann, Katrin and Saladi, Shyam M. and Galimidi, Rachel P. and Zhang, Bin and Clemons, Jr., William M. and Miller, III, Thomas F.},
abstractNote = {},
doi = {10.1016/j.celrep.2016.07.042},
journal = {Cell Reports},
number = 8,
volume = 16,
place = {Netherlands},
year = 2016,
month = 8
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1016/j.celrep.2016.07.042

Save / Share:
  • Specific contacts between the lac repressor and operator have been explored using 5-bromodeoxyuridine-substituted DNA. Substitution of BrdU for single thymidine positions in a synthetic 40-base pair operator provides substrate for ultraviolet irradiation; upon irradiation, strand scission occurs at the BrdU residues. When bound, lac repressor protein provides protection against UV-induced breakage depending on the nature of the sites and type of interaction. We have confirmed 13 unique sites of inducer-sensitive protection along the operator sequence using this method compared to complete substitution with BrdU; differences were observed at two positions for singly substituted versus completely substituted DNAs. The ability ofmore » these photosensitive DNAs to form short range cross-links to bound protein has been used to determine the efficiency with which cross-linked protein-DNA complexes are generated at each individual site of BrdU substitution. Five sites of high efficiency cross-linking to the repressor protein have been identified. At one site, cross-linking without protection from strand scission was observed; this result suggests an unusual mechanism of strand scission and/or cross-linking at this site. Comparison of the UV protection results and the cross-linking data show that these processes provide complementary tools for identifying and analyzing individual protein-DNA contacts.« less
  • The human gene EPB72 coding for the band 7 integral membrane protein, a major protein of the erythrocyte membrane membrane, was isolated from a genomic DNA library and characterized. Spanning {approximately}30 kb, the human EPB72 gene comprises seven exons; intron sizes range from 970 to {approximately}11,200 bp. The first exon contains the 5{prime}-untranslated region (61 nucleotides) and the coding sequence for the N-terminal domain; the second exon encodes the hydrophobic domain, including flanking cysteine and lysine residues. Exon 7 contains the C-terminal portion and a 2-kb 3{prime}-untranslated region. The potential promoter region contains several consensus sequences for ubiquitous transcription factorsmore » (Sp1, AP1, AP2, CP1/2, NF{kappa}B, CREB, Ets-1, and CACC/GT-BF) and two imperfect sequences for erythroid factors (EKLF and GATA-1), in accordance with the ubiquitous distribution of the EPB72 mRNA in different cell types. No TATA box was apparent. An inverted Alu repeat element, flanked by nonamer direct repeats, was identified within the region -913/-620, relative to the cap site. Six additional Alu repeat elements, including one monomer and one trimer, were identified within the introns and the 3{prime}-untranslated region. Two polyadenylation signals in the 3{prime}-noncoding region of exon 7 enable the production of two mRNA species. 45 refs., 3 figs., 2 tabs.« less
  • Bacterial histidine kinases play an important role in the response to external stimuli. Structural studies of the histidine kinase transmembrane domain are challenging due to difficulties in protein expression and sample preparation. After carrying out expression screening of a series of histidine kinases, we investigated sample preparation methods for obtaining high quality samples of the periplasmic and transmembrane domain (PTD) of the bacterial histidine kinase SCO3062. Various sample conditions were tested for their ability to give homogeneous NMR spectra of the SCO3062 PTD with well-resolved resonances. Circular dichroism and 3D {sup 15}N-edited NOESY spectrum results demonstrate that the SCO3062 PTDmore » is predominantly {alpha}-helical. This method should be applicable to the NMR analysis of other transmembrane proteins.« less
  • The authors describe the isolation and characterization of a human gene (CLCN3) and its murine homologue (Clcn3) sharing significant sequence and structural similarities with all previosly identified members of the voltage-gated chloride channel (C1C) family. This gene is expressed primarily in tissues derived from neuroectoderm. Within the brain, Clcn3 expression is particularly evident in the hippocampus, olfactory cortex, and olfactory bulb. CLCN3 encodes a 760-amino-acid protein that differs by only 2 amino acid residues from the protein encoded by Clcn3. CLCN3 protein also shows a high similarity with GEF1, an integral membrane protein of the yeast Saccharomyces cerevisiae known tomore » be involved in respiration and iron-limited cell growth, and with the predicted protein product of a DNA sequence from the mold Septoria nodorum. This high degree of sequence conservation in very distantly related species such as human and yeast indicates that this gene has retained a fundamental function throughout evolution. 42 refs., 6 figs., 1 tab.« less
  • We have isolated and characterized a novel cDNA coding for a highly hydrophobic protein (B5) from a fetal mouse mandibular condyle cDNA library. The full-length mouse B5 cDNA is 3095 nucleotides long and contains a potential open reading frame coding for a protein of 705 amino acids with a calculated molecular weight of 80.5 kDa. The B5 mRNA is differentially polyadenylated, with the most abundant transcript having a length of 2.7 kb. The human homolog of B5 was isolated from a cDNA testis library. The predicted amino acid sequence of the human B5 is 98.5% identical to that of mouse.more » The most striking feature of the B5 protein is the presence of numerous (10-14) potential transmembrane domains, characteristic of an integral membrane protein. Similarity searches in public databanks reveal that B5 is 58% similar to the T12A2.2 gene of Caenorhabditis elegans and 60% similar to the STT3 gene of Saccharomyces cerevisiae. Futhermore, the report of an EST sequence (Accession No. Z13858) related to the human B5, but identical to the STT3 gene, indicates that B5 belongs to a larger gene family coding for novel putative transmembrane proteins. This family exhibits a remarkable degree of conservation in different species. The gene for B5, designated Itm1 (Integral membrane protein 1), is located on mouse chromosome 9. 28 refs., 4 figs.« less