NCCAM Studies

Publication Title: 
Proceedings of the National Academy of Sciences of the United States of America

Mouse immunoglobulin heavy-chain variable region (Ig VH) genes apparently arose from the approximately 600-base-pair-long (approximately 12 tandem repeats of the 48-base-pair-long primordial building block sequence TTC-AGC-AGC-CTG-ACT-GGA-TAT-GAC-CTG-GAG-TGG-ACT-TAC-TGC-GCA-AGA) that in the original reading frame specified the amino acid sequence Phe-Ser-Ser-Leu-Thr-Gly-Tyr-Asp-Leu-Glu-Trp-Thr-Tyr-Cys-Ala-Arg. The previously identified, shorter prototype building blocks merely represented particular portions of the above primordial sequence.

Author(s): 
Ohno, S.
Matsunaga, T.
Wallace, R. B.
Publication Title: 
Journal of Virology

Wild-type Indiana virus transcribed four 11- to 14-nucleotide-long, 5' N-gene mRNA sequences in vitro. The amount of oligonucleotides synthesized relative to leader by wild-type virions varied inversely with the salt concentration of the transcription reaction. Reduced oligonucleotide synthesis by nucleocapsids at all salt concentrations tested and a comparison of the proteins remaining bound to the template of nucleocapsids and virions transcribed in different NaCl concentrations suggested that the matrix (M) protein regulates oligonucleotide synthesis.

Author(s): 
Pinney, D. F.
Emerson, S. U.
Publication Title: 
Immunology

The present studies analysed the uterine free secretory component (SC) response to steroid hormones, and correlated effects on SC with those on IgA. Administration of oestradiol for 3 days to ovariectomized rats significantly increased the levels of SC in uterine secretions, when compared to those in saline-injected controls. This response was dose-dependent and specific for oestrogens, since progesterone, testosterone and glucocorticoids had no effect. The oestradiol-induced elevation in SC levels occurred in parallel with that of IgA.

Author(s): 
Sullivan, D. A.
Underdown, B. J.
Wira, C. R.
Publication Title: 
The Journal of Biological Chemistry

Soluble, truncated mutant and wild-type forms of penicillin-binding protein 5 (sPBP 5) from Escherichia coli were produced in large amounts by placing the dacA gene that encodes PBP 5 under the control of the trp-lac fusion promoter. The 3' end of the dacA gene used in this study contains a stop codon that results in the deletion of 15 amino acids from the carboxyl terminus and the production of a soluble protein. Using oligonucleotide-directed mutagenesis, the role of cysteine 115 in the mechanism of sPBP 5 was investigated.

Author(s): 
Nicholas, R. A.
Strominger, J. L.
Publication Title: 
The Journal of Biological Chemistry

The present study investigates the unique contribution of the NH2-terminal 33 residues of prothrombin, the gamma-carboxyglutamic acid (Gla) domain, to the Ca(II) and phospholipid-binding properties of prothrombin. Two Gla domain peptides, 1-42 and 1-45, produced by chymotryptic cleavage of prothrombin fragment 1 (residues 1-156 of the amino terminus of bovine prothrombin) and isolated by anion-exchange chromatography were utilized to characterize the Gla domain of prothrombin. This investigation utilized several experimental approaches to examine the properties of the Gla domain peptides.

Author(s): 
Pollock, J. S.
Shepard, A. J.
Weber, D. J.
Olson, D. L.
Klapper, D. G.
Pedersen, L. G.
Hiskey, R. G.
Publication Title: 
Proceedings of the National Academy of Sciences of the United States of America

Two VLA proteins (or beta 1 integrins; originally called very late activation antigens) that bind to distinct determinants on fibronectin (FN) are increased on activated immune or memory T cells. VLA-4 binds to the peptide sequence Gly-Pro-Glu-Ile-Leu-Asp-Val-Pro-Ser-Thr (GPEILDVPST in single-letter code) on the alternatively spliced CS-1 form of FN, whereas VLA-5 binds to an Arg-Gly-Asp sequence found on all forms of FN.

Author(s): 
Ferguson, T. A.
Mizutani, H.
Kupper, T. S.
Publication Title: 
Virology

We report the construction of a poliovirus genome [pPVM-VPg(3F4A)] harboring a double mutation in VPg. This mutant, in which the tyrosine and the threonine at residues 3 and 4 of the VPg region were replaced by phenylalanine and alanine, respectively, is lethal, that is, all RNA synthesis was abolished and no revertants could be isolated.

Author(s): 
Cao, X.
Wimmer, E.
Publication Title: 
The EMBO journal

Amongst the picornaviruses, poliovirus encodes a single copy of the genome-linked protein, VPg wheras foot-and-mouth disease virus uniquely encodes three copies of VPg. We have previously shown that a genetically engineered poliovirus genome containing two tandemly arranged VPgs is quasi-infectious (qi) that, upon genome replication, inadvertently deleted one complete VPg sequence. Using two genetically marked viral genomes with two VPg sequences, we now provide evidence that this deletion occurs via homologous recombination.

Author(s): 
Cao, X.
Wimmer, E.
Publication Title: 
Blood

CD34+ precursors in normal human bone marrow (BM) generate large numbers of dendritic cells alongside macrophages and granulocytic precursors when cultured for 12 to 14 days in c-kit ligand, granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha). This study reports an intermediate cell type that develops by day 6, and has the potential to differentiate into either macrophages or dendritic cells.

Author(s): 
Szabolcs, P.
Avigan, D.
Gezelter, S.
Ciocon, D. H.
Moore, M. A.
Steinman, R. M.
Young, J. W.
Publication Title: 
Journal of Immunology (Baltimore, Md.: 1950)

Recent studies have reported that APC can present particulate exogenous Ag in the context of class I MHC to CD8+ CTL, and our laboratory demonstrated that IL-3 could enhance CTL generation to exogenous Ag. In this paper, we wished to determine whether presentation of particulate Ag could be enhanced by IL-3. A T cell hybridoma, B3Z86/90.14 (B3Z) restricted to Ova/Kb, was used as an indicator for presentation of particulate Ag with class I MHC. When activated, this hybridoma expresses lacZ, allowing a simple colorimetric measurement of Ag-specific T cell stimulation.

Author(s): 
Yeh, K. Y.
McAdam, A. J.
Pulaski, B. A.
Shastri, N.
Frelinger, J. G.
Lord, E. M.

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