Prokaryotic Heme Biosynthesis: Multiple Pathways to a Common Essential Product

The advent of heme during evolution allowed organisms possessing this compound to safely and efficiently carry out a variety of chemical reactions that otherwise were difficult or impossible. While it was long assumed that a single heme biosynthetic pathway existed in nature, over the past decade, it has become clear that there are three distinct pathways among prokaryotes, although all three pathways utilize a common initial core of three enzymes to produce the intermediate uroporphyrinogen III. The most ancient pathway and the only one found in the Archaea converts siroheme to protoheme via an oxygen-independent four-enzyme-step process. Bacteria utilize the initial core pathway but then add one additional common step to produce coproporphyrinogen III. Following this step, Gram-positive organisms oxidize coproporphyrinogen III to coproporphyrin III, insert iron to make coproheme, and finally decarboxylate coproheme to protoheme, whereas Gram-negative bacteria first decarboxylate coproporphyrinogen III to protoporphyrinogen IX and then oxidize this to protoporphyrin IX prior to metal insertion to make protoheme. In order to adapt to oxygen-deficient conditions, two steps in the bacterial pathways have multiple forms to accommodate oxidative reactions in an anaerobic environment. The regulation of these pathways reflects the diversity of bacterial metabolism. This diversity, along with the late recognition that three pathways exist, has significantly slowed advances in this field such that no single organism's heme synthesis pathway regulation is currently completely characterized

A new small regulatory protein, HmuP, modulates haemin acquisition in Sinorhizobium meliloti

Sinorhizobium meliloti has multiple systems for iron acquisition, including the use of haem as an iron source. Haem internalization involves the ShmR haem outer membrane receptor and the hmuTUV locus, which participates in haem transport across the cytoplasmic membrane. Previous studies have demonstrated that expression of the shmR gene is negatively regulated by iron through RirA. Here, we identify hmuP in a genetic screen for mutants that displayed aberrant control of shmR. The normal induction of shmR in response to iron limitation was lost in the hmuP mutant, showing that this gene positively affects shmR expression. Moreover, the HmuP protein is not part of the haemin transporter system. Analysis of gene expression and siderophore production indicates that disruption of hmuP does not affect other genes related to the iron-restriction response. Our results strongly indicate that the main function of HmuP is the transcriptional regulation of shmR. Sequence alignment of HmuP homologues and comparison with the NMR structure of Rhodopseudomonas palustris CGA009 HmuP protein revealed that certain amino acids localized within predicted β-sheets are well conserved. Our data indicate that at least one of the β-sheets is important for HmuP activity

Complexity on Small Scales III: Iron and alpha Element Abundances in the Carina Dwarf Spheroidal Galaxy

We have obtained high-resolution spectroscopy of ten red giants in the Carina dwarf spheroidal (dSph) with UVES at the ESO/VLT. Here we present the abundances of O,Na,Mg,Si,Ca,Ti and Fe. By comparing the iron abundances [Fe/H] with calcium triplet (CaT) metallicities we show that the empirical CaT technique yields good agreement with the high-resolution data for [Fe/H]>-2 dex, but tends to deviate at lower metallicities. We identify two metal poor stars with iron abundances of -2.72 and -2.50 dex. These stars are found to have enhanced [alpha/Fe] ratios similar to those of stars in the Milky Way halo. However, the bulk of the Carina red giants are depleted in the [alpha/Fe] abundance ratios with respect to the Galactic halo at a given metallicity. One of our targets, with a [Fe/H] of -1.5 dex, is considerably depleted in almost all of the alpha-elements by ~0.5 dex compared to the solar values. Such a low [alpha/Fe] can be produced by stochastical fluctuations in terms of an incomplete mixing of single Type Ia and II SNe events into the ISM. Our derived element ratios are consistent with the episodic and extended SF in Carina known from its color-magnitude diagram. We find a considerable star-to-star scatter in the abundance ratios. This suggests that Carina's SF history varies with position within the galaxy, with incomplete mixing. Alternatively, the SF rate is so low that the high-mass stellar IMF is sparsely populated, as statistically expected in low-mass star clusters, leading to real scatter in the resultant mass-integrated yields. Both ideas are consistent with slow stochastic SF in dissolving associations, so that one may not speak of a single SF history at a detailed level (Abridged).Comment: 23 pages, 8 figures, accepted for publication in the A

Synthetic Lethality of the bfr and mbfA Genes Reveals a Functional Relationship between Iron Storage and Iron Export in Managing Stress Responses in Bradyrhizobium japonicum.

An mbfA mutant of Bradyrhizobium japonicum defective in iron export is sensitive to short term exposure to high levels iron or H2O2. Here, we found that the mbfA strain grown in elevated iron media (100 μM) became resistant to those treatments, suggesting a stress response adaptation. The bfr gene encodes the iron storage protein bacterioferritin, and its expression is derepressed by iron. An mbfA bfr double mutant showed a loss of stress adaptation, and had a severe growth phenotype in high iron media. Moreover, a bfrup allele in which bfr is constitutively derepressed conferred stress tolerance on an mbfA mutant without elevating the iron content in the growth media. The intracellular iron content of the mbfA bfr double mutant was substantially higher than that found in the wild type, even when grown in relatively low iron media (5 μM). Under that condition, iron-responsive gene expression was aberrant in the mbfA bfr strain. Moreover, the double mutant was sensitive to the iron-activated antibiotic streptonigrin. We conclude that MbfA and Bfr work in concert to manage iron and oxidative stresses. In addition, the need for iron detoxification is not limited to extreme environments, but is also required for normal cellular function