Inactivation of the monocistronic rca gene in Anabaena variabilis suggests a physiological ribulose bisphosphate carboxylase oxygenase activase-like function in heterocystous cyanobacteria

被引:26
作者
Li, LA
Zianni, MR
Tabita, FR
机构
[1] Ohio State Univ, Dept Microbiol, Columbus, OH 43210 USA
[2] Ohio State Univ, Plant Mol Biol Biotechnol Program, Columbus, OH 43210 USA
关键词
activase; CO2; fixation; cyanobacteria; rca gene; Rubisco;
D O I
10.1023/A:1006251808625
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
There was no discernible effect after incubating recombinant Anabaena Rubisco and carboxyarabinitol 1-phosphate with the product of the Anabaena rca gene. Since the unactivated cyanobacterial Rubisco is not readily inhibited by ribulose 1,5-bisphosphate and fallover is not observed, a genetic basis for the function of the Rubisco activase-like gene (rca) was sought. The monocistronic rca gene was inactivated in vivo and resulting mutant strains of A. variabilis were found to be incapable of synthesizing immunologically detected RCA protein. The requirement for the product of the rca gene in the light was further examined by measuring Rubisco activity in permeabilized whole cells of wild-type and rca mutant strains at different light intensities. In a 1% CO2-air atmosphere, inactivation of rca reduced the ability of A. variabilis to elevate Rubisco activity under high light (73 mu mol quanta m(-2) s(-1)), but had little effect under low light (8 mu mol m(-2) s(-1)). For air-grown cultures, differences in the rates exhibited by the wild-type and rca mutant to fully activate Rubisco during a whole-cell assay were enhanced by increases in light intensity. The significance of the rca mutation was underlined by effects on growth as, unlike the wild-type, growth rates did not increase after cells transferred from low to high light intensities. Higher exogenous CO2 concentrations (1%) were required to sustain a normal growth rate for the A. variabilis rca mutant. When grown in air levels of CO2, the rca mutant not only needed longer times to double in cell density but also exhibited greatly diminished Rubisco activity compared with the wild-type strain. Despite the unusual properties of cyanobacterial Rubisco, these results suggest a physiological role for the product of the rca gene in maximizing the activity of Rubisco in heterocystous cyanobacteria.
引用
收藏
页码:467 / 478
页数:12
相关论文
共 56 条
[31]   INVITRO INSERTIONAL MUTAGENESIS WITH A SELECTABLE DNA FRAGMENT [J].
PRENTKI, P ;
KRISCH, HM .
GENE, 1984, 29 (03) :303-313
[32]   ASSOCIATION OF CARBONIC-ANHYDRASE ACTIVITY WITH CARBOXYSOMES ISOLATED FROM THE CYANOBACTERIUM SYNECHOCOCCUS PCC7942 [J].
PRICE, GD ;
COLEMAN, JR ;
BADGER, MR .
PLANT PHYSIOLOGY, 1992, 100 (02) :784-793
[33]   EVIDENCE FOR THE ROLE OF CARBOXYSOMES IN THE CYANOBACTERIAL CO2-CONCENTRATING MECHANISM [J].
PRICE, GD ;
BADGER, MR .
CANADIAN JOURNAL OF BOTANY-REVUE CANADIENNE DE BOTANIQUE, 1991, 69 (05) :963-973
[34]   NEW AND VERSATILE CLONING VECTORS WITH KANAMYCIN-RESISTANCE MARKER [J].
PRIDMORE, RD .
GENE, 1987, 56 (2-3) :309-312
[35]   HIGH SUBSTRATE-SPECIFICITY FACTOR RIBULOSE-BISPHOSPHATE CARBOXYLASE OXYGENASE FROM EUKARYOTIC MARINE-ALGAE AND PROPERTIES OF RECOMBINANT CYANOBACTERIAL RUBISCO CONTAINING ALGAL RESIDUE MODIFICATIONS [J].
READ, BA ;
TABITA, FR .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1994, 312 (01) :210-218
[36]   AMINO-ACID SUBSTITUTIONS IN THE SMALL SUBUNIT OF RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE OXYGENASE THAT INFLUENCE CATALYTIC ACTIVITY OF THE HOLOENZYME [J].
READ, BA ;
TABITA, FR .
BIOCHEMISTRY, 1992, 31 (02) :519-525
[37]   GENERIC ASSIGNMENTS, STRAIN HISTORIES AND PROPERTIES OF PURE CULTURES OF CYANOBACTERIA [J].
RIPPKA, R ;
DERUELLES, J ;
WATERBURY, JB ;
HERDMAN, M ;
STANIER, RY .
JOURNAL OF GENERAL MICROBIOLOGY, 1979, 111 (MAR) :1-61
[38]  
Robinson NJ, 1989, J APPL PHYCOL, V1, P5, DOI 10.1007/bf00003530
[39]   RELEASE OF THE NOCTURNAL INHIBITOR, CARBOXYARABINITOL-1-PHOSPHATE, FROM RIBULOSE BISPHOSPHATE CARBOXYLASE OXYGENASE BY RUBISCO ACTIVASE [J].
ROBINSON, SP ;
PORTIS, AR .
FEBS LETTERS, 1988, 233 (02) :413-416
[40]   ADENOSINE-TRIPHOSPHATE HYDROLYSIS BY PURIFIED RUBISCO ACTIVASE [J].
ROBINSON, SP ;
PORTIS, AR .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1989, 268 (01) :93-99