A tethering mechanism for length control in a processive carbohydrate polymerization.
Carbohydrate polymers are the most abundant organic substances on earth. Their degrees of polymerization range from tens to thousands of units, yet polymerases generate the relevant lengths without the aid of a template. To gain insight into template-independent length control, we investigated how the mycobacterial galactofuranosyl-transferase GlfT2 mediates formation of the galactan, a polymer of galactofuranose residues that is an integral part of the cell wall. We show that isolated recombinant GlfT2 can catalyze the synthesis of polymers with degrees of polymerization that are commensurate with values observed in mycobacteria, indicating that length control by GlfT2 is intrinsic. Investigations using synthetic substrates reveal that GlfT2 is processive. The data indicate that GlfT2 controls length by using a substrate tether, which is distal from the site of elongation. The strength of interaction of that tether with the polymerase influences the length of the resultant polymer. Thus, our data identify a mechanism for length control by a template-independent polymerase.
May
John F
JF
Department of Biochemistry, University of Wisconsin, 433 Babcock Drive, Madison, WI 53706, USA.
Splain
Rebecca A
RA
Brotschi
Christine
C
Kiessling
Laura L
LL
D001426:Bacterial Proteins / Q000378:metabolism; D002241:Carbohydrates / Q000737:chemistry*; D005685:Galactans / Q000096:biosynthesis / Q000737:chemistry; D006639:Histidine / Q000378:metabolism; D008958:Models, Molecular; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D009842:Oligopeptides / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014530:Uridine Diphosphate / Q000378:metabolism
D001426:Bacterial Proteins; D002241:Carbohydrates; D005685:Galactans; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011108:Polymers; D011993:Recombinant Fusion Proteins; D006639:Histidine; D014530:Uridine Diphosphate