Characterization of the nifUHD cluster and a new myoglobin-like gene from Nostoc commune UTEX 584

Sequence analysis of the entire 3.5 kb <i>Hind</i>III genomic DNA fragment previously isolated from <i>Nostoc commune</i> UTEX 584 (Defrancesco and Potts 1988), determined the exact locations of the <i>nifU, nifH</i>, and <i>nifD</i> genes and identified two potential stem loop structures, a direct repeat, and an ORF that codes for a protein with a predicted amino acid sequence similar to that of myoglobin from <i>Paramecium caudatum</i>. The <i>N. commune</i> UTEX 584 myoglobin-like protein has a predicted length of 118 amino acids and molecular mass of 12,906 Da. A PCR copy of the gene (<i>glbN</i>) was cloned for overexpression of the protein. The recombinant protein was purified and used for spectral analysis and for the production of polyclonal antisera. Treatment of the recombinant protein with dithionite and CO resulted in spectral shifts characteristic of hemoproteins that bind oxygen. While some of the spectral characteristics are unique to the protein, in general the spectra were more like those of globins than cytochromes. Based on these characteristics and the sequence similarity to the P. caudatum mnyoglobin, we proposed the name cyanoglobin, with the gene designation glbN and the protein designation GlbN. Western analysis of GlbN expression was performed on N. commune UTEX 584 and two species of Anabaena (Anabaena sp. PCC 7120 and Anabaena variabilis). In N. commune UTEX 584 a protein with a molecular mass similar to that predicted for GlbN was detected. This protein was produced in increased amounts under the same growth conditions that resulted in increased production of nitrogenase reductase (the nifH gene product). No proteins of similar size to GlbN were detected in Anabaena sp. PCC 7120 or A. variabilis. A possible function of GlbN may be for oxygen storage, transport, or protection of the nitrogenase system. These functions as well as those of the direct repeat and the potential stem loop structures and their relationship to nitrogen fixation or other physiological processes in N. commune UTEX 584 require further analysis.

Name thesis:degree_name

Ph. D.

Level thesis:degree_level

doctoral

Discipline thesis:degree_discipline

Biochemistry and Nutrition

Department dc:contributor.department

Biochemistry and Nutrition

Grantor dc:publisher

Virginia Tech

Year dc:date.issued

1992

Author dc:creator

• Angeloni, Stephen V.

Chair dc:contributor.committeechair

• Potts, Malcolm

Committee members dc:contributor.committeemember

• Hess, John L.
• Keenan, Thomas W.
• Lacy, George H.
• Larson, Timothy J.

Statement dc:rights

• In Copyright

Licence dc:rights.uri

• http://rightsstatements.org/vocab/InC/1.0/

Language dc:language.iso

en

Dc Identifier Other

etd-02262007-095956

OAI identifier oai:identifier

oai:vtechworks.lib.vt.edu:10919/37418