Difference between revisions of "Part:BBa K1471000"

 
 
(4 intermediate revisions by 2 users not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1471000 short</partinfo>
 
<partinfo>BBa_K1471000 short</partinfo>
  
 
For mercury bioaccumulation.  
 
For mercury bioaccumulation.  
 +
 +
'''Brief description merE'''
 +
 +
Our part is a yeast RBS with an 8KDa CH3Hg or Hg+2 transmembrane bacterial transporter.
 +
 +
'''Biology'''
 +
 +
MerE is a gene is part of the mer operon, a collection of bacterial genes specialized on the tolerance to various compounds of mercury including methylmercury. It is naturally found in the transposon Tn21 from the plasmid NR1 Shigella flexneri or MB1 in the case of Bacillus megaterium. The general mechanism of the operon can be observed on the following representation (Das S., Dash H. R., 2012):
 +
 +
Schematic presentation of mer operon in narrow-spectrum Gram-negative mercury-resistant bacteria (Das S., Dash H. R., 2012)
 +
Where organomercury compounds are transported inside the bacteria by merP, merT, mer E and merG, followed by the transformation of organic mercury by merB into its ionic form and the reduction from Hg+2 into volatile Hg0 by mer A with the help of NADPH(Das S., Dash H. R., 2012).
 +
Although the transportation of methylmercury is barely understood, there is evidence that recombinant E. coli and other transformed GRAM negative bacteria are able to accumulate mercury thanks to the transformation of organic mercury into its ionic form(Das S., Dash H. R., 2012).
 +
 +
<html>
 +
<center><img src="http://i1088.photobucket.com/albums/i325/mggo93/imagemerE1.png?t=1414699635" style="width: 500px;"></center> </html>
 +
 +
'''Behaviour'''
 +
 +
merE has been characterized previously on A. thaliana as a potential mercury accumulator and transporter. In Kyono,M., et al (2013)´s study shoot and root growth were observed to become more tolerant in transgenic Arabidopsis compared with controls. As it can be seen on the figure:
 +
 +
<html>
 +
<center><img src="http://i1088.photobucket.com/albums/i325/mggo93/imagemerE2.png?t=1414699634" style="width: 700px;"></center> </html>
 +
 +
==MIT_MAHE 2020==
 +
 +
'''Role of cysteine and histidine residues.'''
 +
 +
In order to investigate the molecular function of merE in the transport of methylmercury, a study was conducted with six merE variants with specific site-directed mutations. By comparison methylmercury uptake by cell with intact merE and mutated MerE, it was demonstrated that the cysteine pairs (Cys28 and Cys30) within the first transmembrane domain of MerE and the histidine residue (His51) on the periplasmic face located between the first and second transmembrane domains were required for MerE mediated CH3Hg(I) transport across the cell membrane. The cysteine pairs especially may play a critical role in the transport of CH3Hg(I) and Hg(II). However, it is still unclear why the merE gene found in the T21 mer operon, confers resistance only to Hg(II) and not CH3Hg(I).     
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 17: Line 44:
 
<partinfo>BBa_K1471000 parameters</partinfo>
 
<partinfo>BBa_K1471000 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
===References===
 +
 +
Das S., Dash H. R., (2012). ''Bioremediation of mercury and the importance of bacterial mer genes''. National Institute of Technology.India: International Biodeterioration & Biodegradation. Volume  75. Pages 207-213
 +
 +
Kiyono M. , et al (2013) ''Increase methylmercury accumulation in Arabidopsis thaliana expressing bacterial broad-spectrum mercury transporter MerE''. Springer. Issue 3; Pages 1-13
 +
 +
Kiyono M.,  Sone Y.,  Nakamura R.,  ''et al'' (2013) ''Role of MerC, MerE, MerF, MerT, and/or MerP in Resistance to Mercurials and the Transport of Mercurials in Escherichia coli''. Biological and Pharmaceutical Bulletin. Volume 36; Issue 11; pages 1835-1841
 +
 +
Sone, Y., Uraguchi, S., Takanezawa, Y., Nakamura, R., Pan-Hou, H., & Kiyono, M. (2017). Cysteine and histidine residues are involved in Escherichia coli Tn21 MerE methylmercury transport. FEBS open bio, 7(12), 1994–1999. https://doi.org/10.1002/2211-5463.12341

Latest revision as of 17:14, 19 October 2020

MerE.

For mercury bioaccumulation.

Brief description merE

Our part is a yeast RBS with an 8KDa CH3Hg or Hg+2 transmembrane bacterial transporter.

Biology

MerE is a gene is part of the mer operon, a collection of bacterial genes specialized on the tolerance to various compounds of mercury including methylmercury. It is naturally found in the transposon Tn21 from the plasmid NR1 Shigella flexneri or MB1 in the case of Bacillus megaterium. The general mechanism of the operon can be observed on the following representation (Das S., Dash H. R., 2012):

Schematic presentation of mer operon in narrow-spectrum Gram-negative mercury-resistant bacteria (Das S., Dash H. R., 2012) Where organomercury compounds are transported inside the bacteria by merP, merT, mer E and merG, followed by the transformation of organic mercury by merB into its ionic form and the reduction from Hg+2 into volatile Hg0 by mer A with the help of NADPH(Das S., Dash H. R., 2012). Although the transportation of methylmercury is barely understood, there is evidence that recombinant E. coli and other transformed GRAM negative bacteria are able to accumulate mercury thanks to the transformation of organic mercury into its ionic form(Das S., Dash H. R., 2012).

Behaviour

merE has been characterized previously on A. thaliana as a potential mercury accumulator and transporter. In Kyono,M., et al (2013)´s study shoot and root growth were observed to become more tolerant in transgenic Arabidopsis compared with controls. As it can be seen on the figure:

MIT_MAHE 2020

Role of cysteine and histidine residues.

In order to investigate the molecular function of merE in the transport of methylmercury, a study was conducted with six merE variants with specific site-directed mutations. By comparison methylmercury uptake by cell with intact merE and mutated MerE, it was demonstrated that the cysteine pairs (Cys28 and Cys30) within the first transmembrane domain of MerE and the histidine residue (His51) on the periplasmic face located between the first and second transmembrane domains were required for MerE mediated CH3Hg(I) transport across the cell membrane. The cysteine pairs especially may play a critical role in the transport of CH3Hg(I) and Hg(II). However, it is still unclear why the merE gene found in the T21 mer operon, confers resistance only to Hg(II) and not CH3Hg(I).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

Das S., Dash H. R., (2012). Bioremediation of mercury and the importance of bacterial mer genes. National Institute of Technology.India: International Biodeterioration & Biodegradation. Volume 75. Pages 207-213

Kiyono M. , et al (2013) Increase methylmercury accumulation in Arabidopsis thaliana expressing bacterial broad-spectrum mercury transporter MerE. Springer. Issue 3; Pages 1-13

Kiyono M., Sone Y., Nakamura R., et al (2013) Role of MerC, MerE, MerF, MerT, and/or MerP in Resistance to Mercurials and the Transport of Mercurials in Escherichia coli. Biological and Pharmaceutical Bulletin. Volume 36; Issue 11; pages 1835-1841

Sone, Y., Uraguchi, S., Takanezawa, Y., Nakamura, R., Pan-Hou, H., & Kiyono, M. (2017). Cysteine and histidine residues are involved in Escherichia coli Tn21 MerE methylmercury transport. FEBS open bio, 7(12), 1994–1999. https://doi.org/10.1002/2211-5463.12341