Distinct forms of the ß subunit of GTP-binding regulatory proteins identified by molecular cloning

Two distinct β subunits of guanine nucleotide-binding regulatory proteins have been identified by cDNA cloning and are referred to as β 1 and β 2 subunits. The bovine transducin β subunit (β 1) has been cloned previously. We have now isolated and analyzed cDNA clones that encode the β 2 subunit from bovine adrenal, bovine brain, and a human myeloid leukemia cell line, HL-60. The 340-residue Mr 37,329 β 2 protein is 90% identical with β 1 in predicted amino acid sequence, and it is also organized as a series of repetitive homologous segments. The major mRNA that encodes the bovine β 2 subunit is 1.7 kilobases in length. It is expreβed at lower levels than β 1 subunit mRNA in all tiβues examined. The β 1 and β 2 meβages are expreβed in cloned human cell lines. Hybridization of cDNA probes to bovine DNA showed that β 1 and β 2 are encoded by separate genes. The amino acid sequences for the bovine and human β 2 subunit are identical, as are the amino acid sequences for the bovine and human β 1 subunit. This evolutionary conservation suggests that the two β subunits have different roles in the signal transduction process

Gz, a guanine nucleotide-binding protein with unique biochemical properties

Cloning of a complementary DNA (cDNA) for Gz alpha, a newly appreciated member of the family of guanine nucleotide-binding regulatory proteins (G proteins), has allowed preparation of specific antisera to identify the protein in tissues and to assay it during purification from bovine brain. Additionally, expression of the cDNA in Escherichia coli has resulted in the production and purification of the recombinant protein. Purification of Gz from bovine brain is tedious, and only small quantities of protein have been obtained. The protein copurifies with the beta gamma subunit complex common to other G proteins; another 26- kDa GTP-binding protein is also present in these preparations. The purified protein could not serve as a substrate for NAD-dependent ADP- ribosylation catalyzed by either pertussis toxin or cholera toxin. Purification of recombinant Gz alpha (rGz alpha) from E. coli is simple, and quantities of homogeneous protein sufficient for biochemical analysis are obtained. Purified rGz alpha has several properties that distinguish it from other G protein alpha subunit polypeptides. These include a very slow rate of guanine nucleotide exchange (k = 0.02 min^-1), which is reduced greater than 20-fold in the presence of mM concentrations of Mg2+. In addition, the rate of the intrinsic GTPase activity of Gz alpha is extremely slow. The hydrolysis rate (kcat) for rGz alpha at 30 degrees C is 0.05 min^-1, or 200-fold slower than that determined for other G protein alpha subunits. rGz alpha can interact with bovine brain beta gamma but does not serve as a substrate for ADP-ribosylation catalyzed by either pertussis toxin or cholera toxin. These studies suggest that Gz may play a role in signal transduction pathways that are mechanistically distinct from those controlled by the other members of the G protein family

Repetitive Segmental Structure of the Transducin β Subunit: Homology with the CDC4 Gene and Identification of Related mRNAs

Retinal transducin, a guanine nucleotide regulatory protein (referred to as a G protein) that activates a cGMP phosphodiesterase in photoreceptor cells, is comprised of three subunits. We have identified and analyzed cDNA clones of the bovine transducin β subunit that may be highly conserved or identical to that in other G proteins. From the cDNA nucleotide sequence of the entire coding region, the primary structure of a 340-amino acid protein was deduced. The encoded β subunit has a Mr of 37,375 and is comprised of repetitive homologous segments arranged in tandem. Furthermore, significant homology in primary structure and segmental sequence exists between the β subunit and the yeast CDC4 gene product. The Mr 37,375 β subunit polypeptide is encoded by a 2.9-kilobase (kb) mRNA. However, there exists in retina other β-related mRNAs that are divergent from the 2.9-kb mRNA on the basis of oligonucleotide and primer-extended probe hybridizations. All mammalian tissues and clonal cell lines that have been examined contain at least two β-related mRNAs, usually 1.8 and 2.9 kb in length. These results suggest that the mRNAs are the processed products of a small number of closely related genes or of a single highly complex β gene

Superconductivity-Induced Anomalies in the Spin Excitation Spectra of Underdoped YBa_2 Cu_3 O_{6+x}

Polarized and unpolarized neutron scattering has been used to determine the effect of superconductivity on the magnetic excitation spectra of YBa_2 Cu_3 O_{6.5} (T_c = 52K) and YBa_2 Cu_3 O_{6.7} (T_c = 67K). Pronounced enhancements of the spectral weight centered around 25 meV and 33 meV, respectively, are observed below T_c in both crystals, compensated predominantly by a loss of spectral weight at higher energies. The data provide important clues to the origin of the 40 meV magnetic resonance peak in YBa_2 Cu_3 O_7.Comment: LaTex, 4 pages, 4 ps figures. to appear in Phys. Rev. Let

Generalisations of splicing languages in DNA splicing systems involving two palindromic restriction enzymes

DNA splicing system is initiated by Head to mathematically model a relation between formal language theory and DNA molecules. In DNA splicing systems, DNA molecules are cut and recombined in specific ways with the existence of enzymes, which are also known as endonucleases, to produce further molecules. The resulting molecules are depicted as splicing languages by using concepts in formal languages theory. A sequence of restriction enzyme that reads the same forward and backward is called as a palindromic rule. Previously, researches on different types of splicing languages have been done. In this research, generalisations of splicing languages resulting from DNA splicing systems with non-overlapping cutting sites of two palindromic restriction enzymes are presented as theorems using the induction method. The results from this research are beneficial for researchers in the field of DNA computing since it contributes to the development of splicing languages generated from DNA splicing systems with different palindromic restriction enzymes by using these generalisations

Antiferromagnetism and d-wave superconductivity in cuprates: a uster DMFT study

We present a new approach to investigate the coexistence of antiferromagnetism and d-wave superconductivity in the two dimensional extended Hubbard model within a numerically exact cluster dynamical mean-field approximation. Self-consistent solutions with two non-zero order parameters exists in the wide range of doping level and temperatures. A linearized equation for energy spectrum near the Fermi level have been solved. The resulting d-wave gap has the correct magnitude and k-dependence but some distortion compare to the pure d_{x^2-y^2} superconducting order parameter due to the presence of underlying antiferromagnetic ordering.Comment: 4 pages, 3 figure

Closure properties of bonded sequential insertion-deletion systems

Through the years, formal language theory has evolved through continual interdisciplinary work in theoretical computer science, discrete mathematics and molecular biology. The combination of these areas resulted in the birth of DNA computing. Here, language generating devices that usually considered any set of letters have taken on extra restrictions or modified constructs to simulate the behavior of recombinant DNA. A type of these devices is an insertion-deletion system, where the operations of insertion and deletion of a word have been combined in a single construct. Upon appending integers to both sides of the letters in a word, bonded insertion-deletion systems were introduced to accurately depict chemical bonds in chemical compounds. Previously, it has been shown that bonded sequential insertion-deletion systems could generate up to recursively enumerable languages. However, the closure properties of these systems have yet to be determined. In this paper, it is shown that bonded sequential insertion-deletion systems are closed under union, concatenation, concatenation closure, λ-free concatenation closure, substitution and intersection with regular languages. Hence, the family of languages generated by bonded sequential insertion-deletion systems is shown to be a full abstract family of languages

Pi excitation of the t-J model

In this paper, we present analytical and numerical calculations of the pi resonance in the t-J model. We show in detail how the pi resonance in the particle-particle channel couples to and appears in the dynamical spin correlation function in a superconducting state. The contribution of the pi resonance to the spin excitation spectrum can be estimated from general model-independent sum rules, and it agrees with our detailed calculations. The results are in overall agreement with the exact diagonalization studies of the t-J model. Earlier calculations predicted the correct doping dependence of the neutron resonance peak in the YBCO superconductor, and in this paper detailed energy and momentum dependence of the spin correlation function is presented. The microscopic equations of motion obtained within current formalism agree with that of the SO(5) nonlinear sigma model, where the pi resonance is interpreted as a pseudo Goldstone mode of the spontaneous SO(5) symmetry breaking.Comment: 33 pages, LATEX, 14 eps fig

Aggregatibacter Actinomycetemcomitans Leukotoxin is Post-Translationally Modified by Addition of Either Saturated or Hydroxylated Fatty Acyl Chains

Aggregatibacter actinomycetemcomitans, a common inhabitant of the human upper aerodigestive tract, produces a repeat in toxin (RTX), leukotoxin (LtxA). The LtxA is transcribed as a 114-kDa inactive protoxin with activation being achieved by attachment of short chain fatty acyl groups to internal lysine residues. Methyl esters of LtxA that were isolated from A. actinomycetemcomitans strains JP2 and HK1651 and subjected to gas chromatography/mass spectrometry contained palmitoyl (C16:0, 27–29%) and palmitolyl (C16:1 cis Δ9, 43–44%) fatty acyl groups with smaller quantities of myristic (C14:0, 14%) and stearic (C18:0, 12–14%) fatty acids. Liquid chromatography/mass spectrometry of tryptic peptides from acylated and unacylated recombinant LtxA confirmed that Lys562 and Lys687 are the sites of acyl group attachment. During analysis of recombinant LtxA peptides, we observed peptide spectra that were not observed as part of the RTX acylation schemes of either Escherichia coli α-hemolysin or Bordetella pertussis cyclolysin. Mass calculations of these spectra suggested that LtxA was also modified by the addition of monohydroxylated forms of C14 and C16 acyl groups. Multiple reaction monitoring mass spectrometry identified hydroxymyristic and hydroxypalmitic acids in wild-type LtxA methyl esters. Single or tandem replacement of Lys562 and Lys687 with Arg blocks acylation, resulting in a \u3e75% decrease in cytotoxicity when compared with wild-type toxin, suggesting that these posttranslational modifications are playing a critical role in LtxA-mediated target cell cytotoxicity