The gene of an archaeal α-L-fucosidase is expressed by translational frameshifting

被引:22
作者
Cobucci-Ponzano, Beatrice
Conte, Fiorella
Benelli, Dario
Londei, Paola
Flagiello, Angela
Monti, Maria
Pucci, Piero
Rossi, Mose
Moracci, Marco
机构
[1] CNR, Inst Prot Biochem, I-80131 Naples, Italy
[2] Univ Bari, Dipartimento Biochim Med & Biol Med, DIBIM, I-70124 Bari, Italy
[3] Univ Naples Federico II, CEINGE, Biotecnol Avanzate Scarl, Dipartimento Chim Organ & Biochim, I-80126 Naples, Italy
[4] Univ Naples Federico II, Dipartimento Biol Strutturale & Funz, Complesso Univ Monte S Angelo, I-80126 Naples, Italy
关键词
D O I
10.1093/nar/gkl574
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
The standard rules of genetic translational decoding are altered in specific genes by different events that are globally termed recoding. In Archaea recoding has been unequivocally determined so far only for termination codon readthrough events. We study here the mechanism of expression of a gene encoding for a alpha-L-fucosidase from the archaeon Sulfolobus solfataricus (fucA1), which is split in two open reading frames separated by a -1 frameshifting. The expression in Escherichia coli of the wild-type split gene led to the production by frameshifting of full-length polypeptides with an efficiency of 5%. Mutations in the regulatory site where the shift takes place demonstrate that the expression in vivo occurs in a programmed way. Further, we identify a full-length product of fucA1 in S.solfataricus extracts, which translate this gene in vitro by following programmed -1 frameshifting. This is the first experimental demonstration that this kind of recoding is present in Archaea.
引用
收藏
页码:4258 / 4268
页数:11
相关论文
共 40 条
[1]
P-site tRNA is a crucial initiator of ribosomal frameshifting [J].
Baranov, PV ;
Gesteland, RF ;
Atkins, JF .
RNA, 2004, 10 (02) :221-230
[2]
Release factor 2 frameshifting sites in different bacteria [J].
Baranov, PV ;
Gesteland, RF ;
Atkins, JF .
EMBO REPORTS, 2002, 3 (04) :373-377
[3]
Recoding: translational bifurcations in gene expression [J].
Baranov, PV ;
Gesteland, RF ;
Atkins, JF .
GENE, 2002, 286 (02) :187-201
[4]
Endless possibilities: translation termination and stop codon recognition [J].
Bertram, G ;
Innes, S ;
Minella, O ;
Richardson, JP ;
Stansfield, I .
MICROBIOLOGY-SGM, 2001, 147 :255-269
[5]
Direct charging of tRNACUA with pyrrolysine in vitro and in vivo [J].
Blight, SK ;
Larue, RC ;
Mahapatra, A ;
Longstaff, DG ;
Chang, E ;
Zhao, G ;
Kang, PT ;
Church-Church, KB ;
Chan, MK ;
Krzycki, JA .
NATURE, 2004, 431 (7006) :333-335
[6]
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[7]
Probing the catalytically essential residues of the α-L-fucosidase from the hyperthermophilic Archaeon Sulfolobus solfataricus [J].
Cobucci-Ponzano, B ;
Mazzone, M ;
Rossi, M ;
Moracci, M .
BIOCHEMISTRY, 2005, 44 (16) :6331-6342
[8]
Recoding in Archaea [J].
Cobucci-Ponzano, B ;
Rossi, M ;
Moracci, M .
MOLECULAR MICROBIOLOGY, 2005, 55 (02) :339-348
[9]
Identification of the catalytic nucleophile of the family 29 α-L-fucosidase from Sulfolobus solfataricus via chemical rescue of an inactive mutant [J].
Cobucci-Ponzano, B ;
Trincone, A ;
Giordano, A ;
Rossi, M ;
Moracci, M .
BIOCHEMISTRY, 2003, 42 (32) :9525-9531
[10]
Identification of an archaeal α-L-fucosidase encoded by an interrupted gene -: Production of a functional enzyme by mutations mimicking programmed-1 frameshifting [J].
Cobucci-Ponzano, B ;
Trincone, A ;
Giordano, A ;
Rossi, M ;
Moracci, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (17) :14622-14631