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Journal of Bacteriology, April 1999, p. 2267-2272, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Structure-Function Study of MalF Protein by Random Mutagenesis

María Isabel Tapia, Michaël Mourez, Maurice Hofnung, and Elie Dassa*

Unité de Programmation Moléculaire et Toxicologie Génétique, CNRS URA 1444, Institut Pasteur, F75724 Paris Cedex 15, France

Received 13 October 1998/Accepted 28 January 1999

MalF is one of the two integral inner membrane proteins of the maltose-maltodextrin transport system. To identify functional regions in this protein, we characterized a collection of malF mutants obtained by random mutagenesis. We analyzed their growth on maltose and maltodextrins, the steady-state levels and subcellular localization of the mutant proteins, and the subcellular localization of MalK. Only 2 of the 21 MalF mutant proteins allowed growth on maltose and maltodextrins. Most mutations resulting in immunodetectable proteins mapped to hydrophilic domains, indicating that insertions affecting transmembrane segments gave rise to unstable or lethal proteins. All MalF mutant proteins, even those C-terminally truncated or with large N-terminal deletions, were inserted into the cytoplasmic membrane. Having identified mutations leading to reduced steady-state level, to partial mislocation, and/or to misfolding, we were able to assign to some regions of MalF a role in the assembly of the MalFGK2 complex and/or in the transport mechanism.

* Corresponding author. Mailing address: Unité de Programmation Moléculaire et Toxicologie Génétique, CNRS URA 1444, Institut Pasteur, 25 rue du Dr. Roux, F75724 Paris Cedex 15, France. Phone: 33 (0)1 45 68 88 31. Fax: 33 0(1) 45 68 88 34. E-mail: elidassa{at}pasteur.fr.

Journal of Bacteriology, April 1999, p. 2267-2272, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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