Genes encoding the biotin carboxylase subunit of acetyl-CoA carboxylase from Brassica napus and parental species: cloning, expression patterns, and evolution

Genome ◽  
2011 ◽  
Vol 54 (3) ◽  
pp. 202-211 ◽  
Author(s):  
Zhi-Guo Li ◽  
Wei-Bo Yin ◽  
Li-Ying Song ◽  
Yu-Hong Chen ◽  
Rong-Zhan Guan ◽  
...  
Keyword(s):  
Amino Acid ◽  
Brassica Napus ◽  
Parental Species ◽  
Expression Patterns ◽  
Carrier Protein ◽  
Acetyl Coa Carboxylase ◽  
Transit Peptides ◽  
Genes Encoding ◽  
Group 2 ◽  
Group 1

Comparative genomics is a useful tool to investigate gene and genome evolution. Biotin carboxylase (BC), an important subunit of heteromeric acetyl-CoA carboxylase (ACCase) that is a rate-limiting enzyme in fatty acid biosynthesis in dicots, catalyzes ATP, biotin carboxyl carrier protein, and CO2 to form carboxybiotin carboxyl carrier protein. In this study, we cloned four genes encoding BC from Brassica napus L. (namely BnaC.BC.a, BnaC.BC.b, BnaA.BC.a, and BnaA.BC.b), and two were cloned from each of the two parental species Brassica rapa L. (BraA.BC.a and BraA.BC.b) and Brassica oleracea L. (BolC.BC.a and BolC.BC.b). Sequence analyses revealed that in B. napus the genes BnaC.BC.a and BnaC.BC.b were from the C genome of B. oleracea, whereas BnaA.BC.a and BnaA.BC.b were from the A genome of B. rapa. Comparative and cluster analysis indicated that these genes were divided into two major groups, BnaC.BC.a, BnaA.BC.a, BraA.BC.a, and BolC.BC.a in group-1 and BnaC.BC.b, BnaA.BC.b, BraA.BC.b, and BolC.BC.b in group-2. The divergence of group-1 and group-2 genes occurred in their common ancestor 13–17 million years ago (MYA), soon after the divergence of Arabidopsis and Brassica (15–20 MYA). This time of divergence is identical to the previously reported triplicated time of paralogous subgenomes of diploid Brassica species and the divergence date of group-1 and group-2 genes of α-carboxyltransferase, another subunit of heteromeric ACCase, in Brassica. Reverse transcription PCR revealed that the expression level of group-1 and group-2 genes varied in different organs, and the expression patterns of the two groups of genes were similar in different organs, except in flower. However, two paralogs of group-2 BC genes from B. napus could express differently in mature plants tested by generating BnaA.BC.b and BnaC.BC.b promoter–β-glucuronidase (GUS) fusions. The amino acid sequences of proteins encoded by these genes were highly conserved, except the sequence encoding predicted plastid transit peptides. The plastid transit peptides on the BC precursors of Brassica (71–72 amino acid residues) were predicted based on AtBC protein, compared, and confirmed by fusion with green fluorescent protein. Our results will be helpful in elucidating the evolution and the regulation of ACCase in the genus Brassica.

Genes encoding the α-carboxyltransferase subunit of acetyl-CoA carboxylase from Brassica napus and parental species: cloning, expression patterns, and evolution

Genome ◽  
2010 ◽  
Vol 53 (5) ◽  
pp. 360-370 ◽  
Author(s):  
Zhi-Guo Li ◽  
Wei-Bo Yin ◽  
Huan Guo ◽  
Li-Ying Song ◽  
Yu-Hong Chen ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Fatty Acid Biosynthesis ◽  
Parental Species ◽  
Expression Patterns ◽  
Brassica Species ◽  
Orthologous Gene ◽  
Glucuronidase Activity ◽  
Genes Encoding ◽  
Group 2 ◽  
Group 1

Heteromeric acetyl coenzyme A carboxylase (ACCase), a rate-limiting enzyme in fatty acid biosynthesis in dicots, is a multi-enzyme complex consisting of biotin carboxylase, biotin carboxyl carrier protein, and carboxyltransferase (α-CT and β-CT). In the present study, four genes encoding α-CT were cloned from Brassica napus , and two were cloned from each of the two parental species, B. rapa and B. oleracea . Comparative and cluster analyses indicated that these genes were divided into two major groups. The major divergence between group-1 and group-2 occurred in the second intron. Group-2 α-CT genes represented the ancestral form in the genus Brassica. The divergence of group-1 and group-2 genes occurred in their common ancestor 12.96–17.78 million years ago (MYA), soon after the divergence of Arabidopsis thaliana and Brassica (15–20 MYA). This time of divergence is identical to that reported for the paralogous subgenomes of diploid Brassica species (13–17 MYA). Real-time reverse transcription PCR revealed that the expression patterns of the two groups of genes were similar in different organs, except in leaves. To better understand the regulation and evolution of α-CT genes, promoter regions from two sets of orthologous gene copies from B. napus, B. rapa, and B. oleracea were cloned and compared. The function of the promoter of gene Bnα-CT-1-1 in group-1 and gene Bnα-CT-2-1 in group-2 was examined by assaying β-glucuronidase activity in transgenic A. thaliana. Our results will be helpful in elucidating the evolution and regulation of ACCase in oilseed rape.

scholarly journals Two Auxinic Herbicides Affect Brassica napus Plant Hormone Levels and Induce Molecular Changes in Transcription

Biomolecules ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1153
Author(s):  
Jutta Ludwig-Müller ◽  
Roman Rattunde ◽  
Sabine Rößler ◽  
Katja Liedel ◽  
Freia Benade ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Expression Patterns ◽  
Growth Conditions ◽  
Growth Stages ◽  
Auxin Response ◽  
Time Points ◽  
Genes Encoding ◽  
Different Temperatures ◽  
Indigenous Plant ◽  
Over Time

With the introduction of the new auxinic herbicide halauxifen-methyl into the oilseed rape (Brassica napus) market, there is a need to understand how this new molecule interacts with indigenous plant hormones (e.g., IAA) in terms of crop response. The aim of this study was to investigate the molecular background by using different growth conditions under which three different auxinic herbicides were administered. These were halauxifen-methyl (Hal), alone and together with aminopyralid (AP) as well as picloram (Pic). Three different hormone classes were determined, free and conjugated indole-3-acetic acid (IAA), aminocyclopropane carboxylic acid (ACC) as a precursor for ethylene, and abscisic acid (ABA) at two different temperatures and growth stages as well as over time (2–168 h after treatment). At 15 °C growth temperature, the effect was more pronounced than at 9 °C, and generally, the younger leaves independent of the developmental stage showed a larger effect on the alterations of hormones. IAA and ACC showed reproducible alterations after auxinic herbicide treatments over time, while ABA did not. Finally, a transcriptome analysis after treatment with two auxinic herbicides, Hal and Pic, showed different expression patterns. Hal treatment leads to the upregulation of auxin and hormone responses at 48 h and 96 h. Pic treatment induced the hormone/auxin response already after 2 h, and this continued for the other time points. The more detailed analysis of the auxin response in the datasets indicate a role for GH3 genes and genes encoding auxin efflux proteins. The upregulation of the GH3 genes correlates with the increase in conjugated IAA at the same time points and treatments. Also, genes for were found that confirm the upregulation of the ethylene pathway.

scholarly journals Mamu-B*17+Rhesus Macaques Vaccinated withenv,vif, andnefManifest Early Control of SIVmac239 Replication

2018 ◽  
Vol 92 (16) ◽  
Author(s):  
Mauricio A. Martins ◽  
Damien C. Tully ◽  
Núria Pedreño-Lopez ◽  
Benjamin von Bredow ◽  
Matthias G. Pauthner ◽  
...  
Keyword(s):  
T Cells ◽  
Rhesus Macaques ◽  
Chronic Phase ◽  
Virologic Suppression ◽  
Mhc I ◽  
Immunodeficiency Virus ◽  
Genes Encoding ◽  
Siv Infection ◽  
Group 2 ◽  
Group 1

ABSTRACTCertain major histocompatibility complex class I (MHC-I) alleles are associated with spontaneous control of viral replication in human immunodeficiency virus (HIV)-infected people and simian immunodeficiency virus (SIV)-infected rhesus macaques (RMs). These cases of “elite” control of HIV/SIV replication are often immune-mediated, thereby providing a framework for studying anti-lentiviral immunity. In this study, we examined how vaccination impacts SIV replication in RMs expressing the MHC-I alleleMamu-B*17. Approximately 21% ofMamu-B*17+and 50% ofMamu-B*08+RMs control chronic-phase viremia after SIVmac239 infection. Because CD8+T cells targeting Mamu-B*08-restricted SIV epitopes have been implicated in virologic suppression inMamu-B*08+RMs, we investigated whether this might also be true forMamu-B*17+RMs. Two groups ofMamu-B*17+RMs were vaccinated with genes encoding Mamu-B*17-restricted epitopes in Vif and Nef. These genes were delivered by themselves (group 1) or together withenv(group 2). Group 3 included MHC-I-matched RMs and served as the control group. Surprisingly, the group 1 vaccine regimen had little effect on viral replication compared to group 3, suggesting that unlikeMamu-B*08+RMs, preexisting SIV-specific CD8+T cells alone do not facilitate long-term virologic suppression inMamu-B*17+RMs. Remarkably, however, 5/8 group 2 vaccinees controlled viremia to <15 viral RNA copies/ml soon after infection. No serological neutralizing activity against SIVmac239 was detected in group 2, although vaccine-elicited gp140-binding antibodies correlated inversely with nadir viral loads. Collectively, these data shed new light on the unique mechanism of elite control inMamu-B*17+RMs and implicate vaccine-induced, nonneutralizing anti-Env antibodies in the containment of immunodeficiency virus infection.IMPORTANCEA better understanding of the immune correlates of protection against HIV might facilitate the development of a prophylactic vaccine. Therefore, we investigated simian immunodeficiency virus (SIV) infection outcomes in rhesus macaques expressing the major histocompatibility complex class I alleleMamu-B*17. Approximately 21% ofMamu-B*17+macaques spontaneously controlled chronic phase viremia after SIV infection, an effect that may involve CD8+T cells targeting Mamu-B*17-restricted SIV epitopes. We vaccinatedMamu-B*17+macaques with genes encoding immunodominant epitopes in Vif and Nef alone (group 1) or together withenv(group 2). Although neither vaccine regimen prevented SIV infection, 5/8 group 2 vaccinees controlled viremia to below detection limits shortly after infection. This outcome, which was not observed in group 1, was associated with vaccine-induced, nonneutralizing Env-binding antibodies. Together, these findings suggest a limited contribution of Vif- and Nef-specific CD8+T cells for virologic control inMamu-B*17+macaques and implicate anti-Env antibodies in containment of SIV infection.

scholarly journals Three Homologous Genes Encoding sn-Glycerol-3-Phosphate Acyltransferase 4 Exhibit Different Expression Patterns and Functional Divergence in Brassica napus

2010 ◽  
Vol 155 (2) ◽  
pp. 851-865 ◽  
Author(s):  
Xue Chen ◽  
Martin Truksa ◽  
Crystal L. Snyder ◽  
Aliaa El-Mezawy ◽  
Saleh Shah ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Functional Divergence ◽  
Expression Patterns ◽  
Homologous Genes ◽  
Genes Encoding

Effects of restriction of amino acid supply to the isolated perfused guinea-pig mammary gland

1979 ◽  
Vol 46 (1) ◽  
pp. 69-73 ◽  
Author(s):  
T. Ben Mepham ◽  
Andrew R. Peters ◽  
Stephen Alexandrov
Keyword(s):  
Amino Acids ◽  
Mammary Gland ◽  
Amino Acid ◽  
Guinea Pig ◽  
Essential Amino Acids ◽  
Substrate Solution ◽  
Group 2 ◽  
Group 1

SUMMARYWhen individual essential amino acids were omitted for periods of 40–100 min from the infusate substrate solution in isolated perfused guinea-pig mammary gland experiments, uptake of methionine, tyrosine, phenylalanine, histidine and tryptophan (group 1) was significantly depressed by a mean of 49·8%, whereas the remaining essential amino acids (group 2) showed no significant decrease in uptake. During depletion periods oxidation of [14C\amino acids was increased. The possible significance of the differences in absorption between the 2 groups of amino acids is discussed.

Comparison of Administration of Two Standard Intravenous Amino Acid Formulas to Severely Brain-Injured Patients

1988 ◽  
Vol 22 (10) ◽  
pp. 763-768 ◽  
Author(s):  
Linda G. Ott ◽  
Jack J. Schmidt ◽  
A. Byron Young ◽  
Diana L. Twyman ◽  
Robert P. Rapp ◽  
...  
Keyword(s):  
Amino Acid ◽  
Patient Group ◽  
Nitrogen Balance ◽  
Nitrogen Excretion ◽  
Versus Group ◽  
Significant Difference ◽  
Brain Injured ◽  
Group 2 ◽  
Injured Patients ◽  
Group 1

Twenty severely brain-injured patients with Glasgow Coma Scale scores of 4–9 were prospectively randomized to receive one of two standard amino acid formulas, starting with the first day of hospital admission up to day 14 postinjury. Formula 2 (patient group 2) had 54 percent more leucine, 53 percent more isoleucine, 74 percent more valine, 28 percent less phenylalanine, 31 percent less methionine, 111 percent more proline, 38 percent less alanine, and 38 percent less glycine than formula 1 (patient group 1). Groups 1 and 2 received statistically equal overall mean parenteral nutrition calories and protein (2173 ± 147 vs. 2059 ± 143 kcal, and 77 ± 12 vs. 83.1 ± 6 g, respectively). There was a significant difference in overall mean urinary urea nitrogen excretion (group 1 = 24.6 ± 1.3 vs. group 2= 18.3 ± 1.1, p = 0.02) and nitrogen balance (group 1 = −8.0 ± 2.1 vs. group 2 = + 1.8 ± 1.2, p = 0.01). Mean overall isoleucine values were significantly higher in group 2 (overall mean 77 μmol/L vs. 62 μmol/L, p = 0.04). Phenylalanine levels were significantly higher in group 1 (107 μmol/L) versus group 2 (82 μmol/L) patients (p = 0.01). Arginine levels were significantly higher in group 1 (78 μmol/L) versus group 2 (49 μmol/L) patients (p = 0.0002). This observation suggests that some standard intravenous amino acid formulas may be more apt to promote positive nitrogen balance than others.

scholarly journals Amino acid substitutions can influence the natural killer (NK)-mediated recognition of HLA-C molecules. Role of serine-77 and lysine-80 in the target cell protection from lysis mediated by "group 2" or "group 1" NK clones.

1995 ◽  
Vol 182 (2) ◽  
pp. 605-609 ◽  
Author(s):  
R Biassoni ◽  
M Falco ◽  
A Cambiaggi ◽  
P Costa ◽  
S Verdiani ◽  
...  
Keyword(s):  
Amino Acid ◽  
Natural Killer ◽  
Nk Cell ◽  
Site Directed Mutagenesis ◽  
Single Amino Acid ◽  
Amino Acid Substitutions ◽  
Cell Protection ◽  
Cell Clones ◽  
Group 2 ◽  
Group 1

Natural killer (NK) cells have been shown to express a clonally distributed ability to recognize HLA class I alleles. The previously defined NK clones belonging to "group 1" recognize HLA-C*0401 (Cw4) and other HLA-C alleles sharing Asn at position 77 and Lys at position 80. Conversely, the "group 2" NK clones recognize HLA-Cw*0302 (Cw3) and other HLA-C alleles characterized by Ser at position 77 and Asn at position 80. We assessed directly the involvement of these two residues in the capacity of NK cell clones to discriminate between the two groups of HLA-C alleles. To this end, Cw3 and Cw4 alleles were subjected to site-directed mutagenesis. Substitution of the amino acids typical of the Cw3 allele (Ser-77 and Asn-80) with those present in Cw4 (Asn-77 and Lys-80) resulted in a Cw3 mutant that was no longer recognized by group 2 NK cell clones, but that was recognized by group 1 clones. Analysis of Cw3 or Cw4 molecules containing single amino acid substitutions indicates roles for Lys-80 in recognition mediated by group 1 clones and for Ser-77 in recognition mediated by group 2 clones. These results demonstrate that NK-mediated specific recognition of HLA-C allotypes is affected by single natural amino acid substitutions at positions 77 and 80 of the heavy chain.

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