scholarly journals A human cDNA sequence with homology to non-mammalian lysophosphatidic acid acyltransferases

1997 ◽  
Vol 326 (2) ◽  
pp. 455-461 ◽  
Author(s):  
Alasdair C. STAMPS ◽  
Moira A. ELMORE ◽  
Maxine E. HILL ◽  
Kenneth KELLY ◽  
Ashraff A. MAKDA ◽  
...  
Keyword(s):  
Lysophosphatidic Acid ◽  
Cdna Sequence ◽  
Northern Blot Analysis ◽  
U937 Cell ◽  
Splice Variants ◽  
Fatty Acyl ◽  
Acyltransferase Activity ◽  
E Coli ◽  
Human Cdna Sequence ◽  
Rapid Amplification

A novel human homologue of Escherichia coli, yeast and plant 1-acylglycerol-3-phosphate acyltransferase has been isolated from U937 cell cDNA. Expression of the cloned sequence in 1-acylglycerol-3-phosphate acyltransferase-deficient E. coli resulted in increased incorporation of oleic acid into cellular phospholipids. Membranes made from COS7 cells transfected with the cDNA exhibited higher acyltransferase activity towards a range of donor fatty acyl-CoAs and lysophosphatidic acid. Northern-blot analysis of the cDNA sequence indicated high levels of expression in immune cells and epithelium. Rapid amplification of cDNA ends revealed differentially expressed splice variants, which suggests regulation of the enzyme by alternative splicing. This cDNA therefore represents the first described sequence of a mammalian gene homologous to non-mammalian lysophosphatidic acid acyltransferases.

scholarly journals A Novel Lysophosphatidic Acid Acyltransferase of Escherichia coli Produces Membrane Phospholipids with a cis-vaccenoyl Group and Is Related to Flagellar Formation

Biomolecules ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 745
Author(s):  
Yosuke Toyotake ◽  
Masayoshi Nishiyama ◽  
Fumiaki Yokoyama ◽  
Takuya Ogawa ◽  
Jun Kawamoto ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lysophosphatidic Acid ◽  
Immunoblot Analysis ◽  
Fatty Acyl ◽  
Temperature Sensitive ◽  
Membrane Phospholipids ◽  
E Coli ◽  
Acyl Composition ◽  
Lysophosphatidic Acid Acyltransferase ◽  
Fatty Acyl Composition

Lysophosphatidic acid acyltransferase (LPAAT) introduces fatty acyl groups into the sn-2 position of membrane phospholipids (PLs). Various bacteria produce multiple LPAATs, whereas it is believed that Escherichia coli produces only one essential LPAAT homolog, PlsC—the deletion of which is lethal. However, we found that E. coli possesses another LPAAT homolog named YihG. Here, we show that overexpression of YihG in E. coli carrying a temperature-sensitive mutation in plsC allowed its growth at non-permissive temperatures. Analysis of the fatty acyl composition of PLs from the yihG-deletion mutant (∆yihG) revealed that endogenous YihG introduces the cis-vaccenoyl group into the sn-2 position of PLs. Loss of YihG did not affect cell growth or morphology, but ∆yihG cells swam well in liquid medium in contrast to wild-type cells. Immunoblot analysis showed that FliC was highly expressed in ∆yihG cells, and this phenotype was suppressed by expression of recombinant YihG in ∆yihG cells. Transmission electron microscopy confirmed that the flagellar structure was observed only in ∆yihG cells. These results suggest that YihG has specific functions related to flagellar formation through modulation of the fatty acyl composition of membrane PLs.

Identification of two lysophosphatidic acid acyltransferase genes with overlapping function in Pseudomonas fluorescens

Microbiology ◽  
2005 ◽  
Vol 151 (9) ◽  
pp. 3071-3080 ◽  
Author(s):  
Méabh Cullinane ◽  
Christine Baysse ◽  
John P. Morrissey ◽  
Fergal O'Gara
Keyword(s):  
Pseudomonas Fluorescens ◽  
Lysophosphatidic Acid ◽  
Elevated Temperatures ◽  
Acid Methyl Ester ◽  
Acyltransferase Activity ◽  
E Coli ◽  
Growth Defects ◽  
Acid Methyl ◽  
Identification And Characterization

Phosphatidic acid (PA) is known to be a crucial phospholipid intermediate in cell membrane biosynthesis. In Escherichia coli, this molecule is produced from lysophosphatidic acid (LPA) by LPA acyltransferase (EC 2.3.1.51), encoded by plsC. E. coli possesses only one such LPA acyltransferase and a plsC mutant is non-permissive for growth at elevated temperatures. This study describes the identification and characterization of two genes from Pseudomonas fluorescens F113 that encode enzymes with LPA acyltransferase activity. One of the genes, hdtS, was previously described, whereas patB is a novel gene. In addition, a putative lyso-ornithine lipid acyltransferase was also identified. All three proteins possess conserved acyltransferase domains and are homologous to PlsC and to LPA acyltransferases identified in Neisseria meningitidis. Functional analysis determined that both HdtS and PatB are functional LPA acyltransferases, as both complemented an E. coli plsC mutant. Mutants lacking each of the putative acyltransferases were constructed and analysed. Growth defects were observed for hdtS and patB single mutants, and a double hdtSpatB mutant could not be constructed. To determine precise roles in phospholipid synthesis, fatty acid methyl ester analysis was carried out. The hdtS mutant displayed a profile consistent with a defect in LPA acyltransferase activity, whereas no such phenotype was observed in the patB mutant, indicating that hdtS encodes the primary LPA acyltransferase in the cell. The presence of at least two genes specifying LPA acyltransferase activity may have implications for the function and survival of P. fluorescens in diverse environments.

scholarly journals Chromosomal protein HMG-14. Complete human cDNA sequence and evidence for a multigene family.

1986 ◽  
Vol 261 (34) ◽  
pp. 16082-16086 ◽  
Author(s):  
D Landsman ◽  
T Srikantha ◽  
R Westermann ◽  
M Bustin
Keyword(s):  
Multigene Family ◽  
Cdna Sequence ◽  
Chromosomal Protein ◽  
Human Cdna ◽  
Human Cdna Sequence

scholarly journals Synthesis and glycosylation of CD52, the major 'maturation-associated' antigen of rat spermatozoa, in the cauda epididymidis

Reproduction ◽  
2001 ◽  
pp. 435-446 ◽  
Author(s):  
P Derr ◽  
CH Yeung ◽  
TG Cooper ◽  
C Kirchhoff
Keyword(s):  
Blot Analysis ◽  
Cdna Sequence ◽  
Northern Blot Analysis ◽  
Lectin Binding ◽  
Transcriptional Level ◽  
Lectin Blot ◽  
Molecular Masses ◽  
Rat Epididymis ◽  
Sperm Membrane ◽  
Cauda Epididymidis

A western and lectin blot analysis was performed of the major 'maturation-associated' antigen of rat spermatozoa, which is the rat counterpart of human CD52. In the absence of a suitable antibody, direct study of this approximately 26 kDa antigen, named previously SMemG, had been difficult. In the present study, these problems were overcome by raising a polyclonal antibody against a chemosynthetic peptide predicted from the cDNA sequence of the antigen. The antibody bound to a glycoprotein of rat cauda epididymidal tissue and spermatozoa, this glycoprotein was cleaved by phosphatidylinositol-specific phospholipase C and, after deglycosylation, was reduced to approximately 6 kDa. Northern blot analysis confirmed that the CD52 mRNA was transcribed only post-testicularly, and antibody binding to testicular and sperm proteins of different molecular masses was shown to be nonspecific. Flow cytometry also indicated that the antigen was inserted into the sperm membrane during epididymal transit. Moreover, despite the presence of CD52 mRNA in all parts of the rat epididymis, only the 'long' mRNA molecules of the cauda region were efficiently translated and the antigen glycosylated, indicating that expression of rat CD52 is regulated on a post-transcriptional level. Lectin binding and deglycosylation studies supported the contention that there is extensive mucin-type O-glycosylation of rat CD52. In rats, there was no indication of complex N-linked carbohydrates similar to those described for human CD52.

scholarly journals Differential expression of Zymomonas mobilis sucrase genes (sacB and sacC) in Escherichia coli and sucrase mutants of Zymomonas mobilis

2004 ◽  
Vol 47 (3) ◽  
pp. 329-338 ◽  
Author(s):  
Sangiliyandi Gurunathan ◽  
Paramasamy Gunasekaran
Keyword(s):  
Copy Number ◽  
Zymomonas Mobilis ◽  
Northern Blot Analysis ◽  
Shuttle Vector ◽  
Transcript Levels ◽  
E Coli ◽  
Low Copy Number ◽  
Genes Encoding ◽  
Sacb Gene ◽  
In Cells

The sacB and sacC genes encoding levansucrase and extracellular sucrase respectively were independently subcloned in pBluescript (high copy number) and in Z. mobilis-E. coli shuttle vector, pZA22 (low copy number). The expression of these genes were compared under identical background of E. coli and Z. mobilis host. The level of sacB gene expression in E. coli was almost ten fold less than the expression of sacC gene, irrespective of the growth medium or the host strain. In Z. mobilis the expression of sacB and sacC genes was shown to be subject to carbon source dependent regulation. The transcript of sacB and sacC was three fold higher in cells grown on sucrose than in cells grown on glucose/fructose. Northern blot analysis revealed that the transcript levels of sacC was approximately 2-3 times higher than that of sacB. These results suggested that the expression of sacC gene was more pronounced than sacB.

scholarly journals Organization and sequence of the gene for the human mitochondrial dicarboxylate carrier: evolution of the carrier family

1999 ◽  
Vol 344 (3) ◽  
pp. 953-960 ◽  
Author(s):  
Giuseppe FIERMONTE ◽  
Vincenza DOLCE ◽  
Roberto ARRIGONI ◽  
Michael J. RUNSWICK ◽  
John E. WALKER ◽  
...  
Keyword(s):  
Mitochondrial Membrane ◽  
Cdna Sequence ◽  
Gene Sequence ◽  
Alu Sequences ◽  
Human Cdna ◽  
Mitochondrial Inner Membrane ◽  
Human Dna ◽  
Human Genes ◽  
Human Cdna Sequence ◽  
Liver And Kidney

The dicarboxylate carrier (DIC) is a nuclear-encoded protein located in the mitochondrial inner membrane. It catalyses the transport of dicarboxylates such as malate and succinate across the mitochondrial membrane in exchange for phosphate, sulphate and thiosulphate. We have determined the sequences of the human cDNA and gene for the DIC. The gene sequence was established from overlapping genomic clones generated by PCRs by use of primers and probes based upon the human cDNA sequence. It is spread over 8.6 kb of human DNA and is divided into 11 exons. Five short interspersed repetitive Alu sequences are found in intron I. The protein encoded by the gene is 287 amino acids long. In common with the rat protein, it does not have a processed presequence to help to target it into mitochondria. It has been demonstrated by Northern- and Western-blot analyses that the DIC is present in high amounts in liver and kidney, and at lower levels in all the other tissues analysed. The positions of introns contribute towards an understanding of the processes involved in the evolution of human genes for carrier proteins.

scholarly journals Acyltransferase-mediated selection of the length of the fatty acyl chain and of the acylation site governs activation of bacterial RTX toxins

2020 ◽  
Vol 295 (28) ◽  
pp. 9268-9280 ◽  
Author(s):  
Adriana Osickova ◽  
Humaira Khaliq ◽  
Jiri Masin ◽  
David Jurnecka ◽  
Anna Sukova ◽  
...  
Keyword(s):  
Acyl Chain ◽  
Fatty Acyl ◽  
Biologically Active ◽  
Fatty Acyl Chain ◽  
Acyl Carrier Proteins ◽  
E Coli ◽  
Rtx Toxins ◽  
Wide Range ◽  
Lysine Residues ◽  
Acyl Chains

In a wide range of organisms, from bacteria to humans, numerous proteins have to be posttranslationally acylated to become biologically active. Bacterial repeats in toxin (RTX) cytolysins form a prominent group of proteins that are synthesized as inactive protoxins and undergo posttranslational acylation on ε-amino groups of two internal conserved lysine residues by co-expressed toxin-activating acyltransferases. Here, we investigated how the chemical nature, position, and number of bound acyl chains govern the activities of Bordetella pertussis adenylate cyclase toxin (CyaA), Escherichia coli α-hemolysin (HlyA), and Kingella kingae cytotoxin (RtxA). We found that the three protoxins are acylated in the same E. coli cell background by each of the CyaC, HlyC, and RtxC acyltransferases. We also noted that the acyltransferase selects from the bacterial pool of acyl–acyl carrier proteins (ACPs) an acyl chain of a specific length for covalent linkage to the protoxin. The acyltransferase also selects whether both or only one of two conserved lysine residues of the protoxin will be posttranslationally acylated. Functional assays revealed that RtxA has to be modified by 14-carbon fatty acyl chains to be biologically active, that HlyA remains active also when modified by 16-carbon acyl chains, and that CyaA is activated exclusively by 16-carbon acyl chains. These results suggest that the RTX toxin molecules are structurally adapted to the length of the acyl chains used for modification of their acylated lysine residue in the second, more conserved acylation site.

scholarly journals Enhanced Production of Fatty Acid Ethyl Ester with Engineered fabHDG Operon in Escherichia coli

2019 ◽  
Vol 7 (11) ◽  
pp. 552 ◽  
Author(s):  
Ziaur Rahman ◽  
Bong Hyun Sung ◽  
Javed Nawab ◽  
Muhammad Faisal Siddiqui ◽  
Abid Ali ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Ethyl Ester ◽  
Feedback Inhibition ◽  
Acid Ethyl Ester ◽  
Fatty Acid Ethyl Ester ◽  
Fatty Acyl ◽  
Dodecanoic Acid ◽  
E Coli ◽  
Enhanced Production

Biodiesel, or fatty acid ethyl ester (FAEE), is an environmentally safe, next-generation biofuel. Conventionally, FAEE is produced by the conversion of oil/fats, obtained from plants, animals, and microorganisms, by transesterification. Recently, metabolic engineering of bacteria for ready-to-use biodiesel was developed. In Escherichia coli, it is produced by fatty acyl-carrier proteins and ethanol, with the help of thioesterase (TesB) and wax synthase (WS) enzymes. One of the foremost barriers in microbial FAEE production is the feedback inhibition of the fatty acid (FA) operon (fabHDG). Here, we studied the effect of biodiesel biosynthesis in E. coli with an engineered fabHDG operon. With a basic FAEE producing BD1 strain harboring tes and ws genes, biodiesel of 32 mg/L were produced. Optimal FAEE biosynthesis was achieved in the BD2 strain that carries an overexpressed operon (fabH, fabD, and fabG genes) and achieved up to 1291 mg/L of biodiesel, a 40-fold rise compared to the BD1 strain. The composition of FAEE obtained from the BD2 strain was 65% (C10:C2, decanoic acid ethyl ester) and 35% (C12:C2, dodecanoic acid ethyl ester). Our findings indicate that overexpression of the native FA operon, along with FAEE biosynthesis enzymes, improved biodiesel biosynthesis in E. coli.

scholarly journals A human cDNA sequence for a novel glutathione peroxidase-related selenopeptide, GPRP

1989 ◽  
Vol 17 (15) ◽  
pp. 6390-6390 ◽  
Author(s):  
Deborah K. Dunn ◽  
David D. Howells ◽  
Jane P. Richardson ◽  
Peter S. Goldfarb
Keyword(s):  
Glutathione Peroxidase ◽  
Cdna Sequence ◽  
Human Cdna ◽  
Human Cdna Sequence

scholarly journals Ganglioside-binding specificities of E. coli enterotoxin LT-IIc: Importance of long-chain fatty acyl ceramide

Glycobiology ◽  
2012 ◽  
Vol 23 (1) ◽  
pp. 23-31 ◽  
Author(s):  
C. S. Berenson ◽  
H. F. Nawar ◽  
R. L. Kruzel ◽  
L. M. Mandell ◽  
T. D. Connell
Keyword(s):  
Fatty Acyl ◽  
E Coli ◽  
Long Chain
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