Difference between revisions of "Part:BBa K1640006"

Latest revision as of 01:33, 19 September 2015

psbELJ

Sequence and Features

Overview

This biobrick contains three genes from the photosystem II complex of Chlamydomonas reinhardtii - psbE, psbL, and psbJ, driven by Plac promoter, with RBS preceeding each gene, and terminator at the 3' end.

Names:

- PsbE: Cytochrome b559 subunit alpha.
- PsbL: Photosystem II reaction centre protein L.
- PsbJ: Photosystem II core complex subunit.

This part forms operon 2 in our set of photosystem II operons:

Usage and Biology

The psbE gene encodes the alpha subunit of cytochrome b559, an essential component of photosystem II. Its transcription results in an mRNA of 0.3 kb (Mor, T. S., et al., 1995). It contains two introns of 326 and 350 bp (Cushman, J. C., et al., 1988). In C. reinhardtii, the gene is transcribed independently. However, it has been observed in all other studied photosynthetic organisms to form a cluster with psbF, psbL and psbJ (Kiss, É., et al., 2012).

The PsbL gene is a component of the photosystem II reaction centre. It encodes a 4.5 kDA protein that has a single membrane-spanning alpha helical domain (Anbudurai, P. R., & Pakrasi, H., B 1993). This protein supports function of the primary quinone acceptor found in the reaction centre core complex (Kitamura, K., et al., 1994). Furthermore, its carboxyl terminal end plays a role in recovering the electron transfer activity in the complex (Ozawa, S., et al., 1997).

The psbJ gene has a molecular weight of 4.1 kDa and features a single transmembrane helix (Nowaczyk, M. M., et al., 2012). Although the protein it encodes is not significantly involved in photochemical processes, it has been found to be responsible for the regulation of the number of photosystem II complexes in the thylakoid membrane (Lind, L. K., et al., 1993). Studies of mutants have shown that organisms without the psbJ gene have lower electron transfer efficiency in the photosystem II complex (Regel, R. E., et al., 2001).

The psbELJ cluster consists of three subunits that, together with psbF, are essential in the function of photosystem II, as well as the maintenance of the general structure of thylakoid membrane complexes.

Protein information

psbE

mass: 9.3kDa

sequence: MAGKPVERPFSDILTSIRYWVIHSITVPALFIAGWLFVSTGLAYDVFGTPRPNEYFTEDRQEAPLITDRFNALEQVKKLSGN

psbL

mass: 4.43kDa

sequence: MARPNPNKQVVELNRTSLYWGLLLIFVLAVLFSSYIFN

psbJ

mass: 4.29kDa

sequence: MSNTGTTGRIPLWLVGTVVGTLAIGLLAVFFYGSYVGLGSSL

Part verification

Visualisation of psbELJ showing expected banding is shown in the following gel image, top row third set of lanes, with left lane part showing EcoRI digest, right lane showing EcoRI + PstI double digest. This part has been sequenced to confirm design with final biobrick.

References

Anbudurai, P. R., & Pakrasi, H. B. (1993). Mutational analysis of the PsbL protein of photosystem II in the cyanobacterium Synechocystis sp. PCC 6803. Zeitschrift für Naturforschung C, 48(3-4), 267-274.

Cushman, J. C., Christopher, D. A., Little, M. C., Hallick, R. B., & Price, C. A. (1988). Organization of the psbE, psbF, orf38, and orf42 gene loci on the Euglena gracilis chloroplast genome. Current genetics, 13(2), 173-180.

Lind, L. K., Shukla, V. K., Nyhus, K. J., & Pakrasi, H. B. (1993). Genetic and immunological analyses of the cyanobacterium Synechocystis sp. PCC 6803 show that the protein encoded by the psbJ gene regulates the number of photosystem II centers in thylakoid membranes. Journal of Biological Chemistry, 268(3), 1575-1579.

Kiss, É., Kós, P. B., Chen, M., & Vass, I. (2012). A unique regulation of the expression of the psbA, psbD, and psbE genes, encoding the D1, D2 and cytochrome b559 subunits of the Photosystem II complex in the chlorophyll d containing cyanobacterium Acaryochloris marina. Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1817(7), 1083-1094.

Kitamura, K., Ozawa, S., Shiina, T., & Toyoshima, Y. (1994). L protein, encoded by psbL, restores normal functioning of the primary quinone acceptor, Q A, in isolated D1/D2/CP47/Cytb-559/I photosystem II reaction center core complex. FEBS letters, 354(1), 113-116.

Nowaczyk, M. M., Krause, K., Mieseler, M., Sczibilanski, A., Ikeuchi, M., & Rögner, M. (2012). Deletion of psbJ leads to accumulation of Psb27–Psb28 photosystem II complexes in Thermosynechococcus elongatus. Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1817(8), 1339-1345.

Ozawa, S., Kobayashi, T., Sugiyama, R., Hoshida, H., Shiina, T., & Toyoshima, Y. (1997). Role of PSII-L protein (psbL gene product) on the electron transfer in photosystem II complex. 1. Over-production of wild-type and mutant versions of PSII-L protein and reconstitution into the PSII core complex. Plant molecular biology, 34(1), 151-161.

Regel, R. E., Ivleva, N. B., Zer, H., Meurer, J., Shestakov, S. V., Herrmann, R. G., ... & Ohad, I. (2001). Deregulation of electron flow within photosystem II in the absence of the PsbJ protein. Journal of Biological Chemistry, 276(44), 41473-41478.