Isolation and characterization of a cDNA clone encoding an 18-kDa hydrophobic photosystem I subunit (PSI-L) from barley (Hordeum vulgare L.).
Photosystem I in barley contains a polypeptide with an apparent molecular mass of 14 kDa. The polypeptide is N-terminally blocked to amino acid sequencing, but partial amino acid sequences have been determined from three fragments obtained by chemical and enzymatic cleavage. Using an oligonucleotide probe specifying this amino acid sequence, a full length cDNA clone was isolated. The deduced amino acid sequence does not correspond to any previously identified photosystem I subunit. We designate the novel photosystem I subunit PSI-L and the corresponding nuclear gene PsaL. The cDNA clone encodes a precursor polypeptide of 209 amino acid residues with a deduced molecular mass of 22,210 Da. The precursor has a transit peptide typical of proteins imported into chloroplasts. Based on a putative maturation site, the deduced molecular mass of the mature protein is 18 kDa. The PSI-L polypeptide is hydrophobic and predicted to have at least two membrane-spanning alpha-helices. Northern blot analysis shows that the expression of the PsaL gene is light-induced similar to other of the barley photosystem I genes. Southern blot analysis indicates that PsaL is a single copy gene. Partial amino acid sequences of an N-terminally blocked 9-kDa polypeptide show high sequence similarity to the PSI-G polypeptide of spinach and Chlamydomonas reinhardtii. The gene product of PsaG in spinach has previously been assigned as subunit V (Steppuhn, J., Hermans, J., Nechushtai, R., Ljungberg, U., Thümmler, F., Lottspeich, F., and Herrmann, R. G. (1988) FEBS Lett. 237, 218-224). The present study suggests that PSI-L is equivalent to subunit V and that PSI-G is a subunit migrating closely to PSI-H (subunit VI) and PSI-C (subunit VII).
Okkels
J S
JS
Department of Plant Biology, Royal Veterinary and Agricultural University, Frederiksberg C, Copenhagen, Denmark.
Scheller
H V
HV
Svendsen
I
I
Møller
B L
BL
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D001467:Hordeum / Q000235:genetics* / Q000378:metabolism; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics*; D045331:Photosystem I Protein Complex; D010944:Plants / Q000235:genetics; D011487:Protein Conformation; D012689:Sequence Homology, Nucleic Acid
D046911:Macromolecular Substances; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D004247:DNA