Download A cDNA Encoding Ribosomal Protein S4e from

Plant Physiol. (1995) 108: 431-432
Plant Gene Register
A c D N A Encoding Ribosomal Protein S4e from Cotton
(Gossypium hirsutum 1.)
Rickie B. Turley*, David 1. Ferguson, and William R. Meredith, Jr.
United States Department of Agriculture, Agricultura1 Research Service, Cotton Physiology and Genetics
Research, P.O. Box 345, Stoneville, Mississippi 38776
Ribosomes are complex ribonucleoprotein structures
that function as the machinery for synthesizing proteins
needed for the maintenance, growth, development, and
reproduction of cells. During the biogenesis of these structures, ribosomal precursors, i.e. ribonucleoprotein particles, are formed by the assembly of rRNA and ribosomal
proteins in the nucleolus. These particles are processed to
mature ribosomes after they traverse the nuclear membrane into the cytoplasm (Perry, 1976).Therefore, ribosome
biogenesis is not only dependent on transcription and
translation but also on the nuclear import of ribosomal
proteins and the nuclear export of ribosomal particles.
Many of these stages are well documented for bacteria,
yeast, and animal systems (Wittmann-Liebold, 1986);however, little documentation exists for plant ribosomes (Gao
et al., 1994). We have sequenced a cDNA encoding ribosomal protein S4e from cotton (Gossypium hirsutum L. cv
Deltapine 62).
A cDNA encoding a partial-length S4e was identified
and selected from a group of approximately 60 cDNAs
[constructed from leaf poly(A)+ RNA] that demonstrated
restriction fragment length polymorphisms with genomic
DNA from G. hivsutum. Sequencing of the cDNAs was used
to deduce the amino acid composition and identify proteins using QGSEARCH and the SwissProt data bases in
PC/Gene (Intelligenetics, Inc., Mountain View, CA).' The
partial-length S4e cDNA was used to screen a Uni-Zap
library constructed by Ni and Trelease (1991).A full-length
cDNA with a 1.09-kb insert was identified, and the nucleotide sequence was determined in its entirety for both
strands (Table I). The clone has a 5' (55 bp) and 3' (249 bp)
noncoding region, with a 786-bp open reading frame. The
deduced amino acid sequence has 262 residues with an M ,
of 29,635. The protein contains 58 basic residues (Arg, His,
and Lys) and only 24 acidic residues (Asp and Glu). No
consensus polyadenylation signal was found in the 3' noncoding region, as is common in many plant "As.
A comparison was made of the deduced amino acid
sequence of the cotton S4e protein with four S4e proteins
Table 1. Nucleotide and deduced amino acid sequence of ribosomal protein S4e in cotton cotyledons
Organism:
Cotton (Gossypium hirsutum L. cv DPL 62).
Function:
40s ribosomal protein S4e.
Clone Designation:
S4e.
Cene Location:
Multiple copy, nuclear gene.
Method of Identification:
Sequence comparison with ribosomal protein S4e from yeast
(65% identical), mouse (70% identical), and human X (69%
identical) and Y (66% identical) chromosomes.
Sequencing Strategy:
Plasmid sequenced using double-stranded templates and
dideoxynucleotide chain termination method.
Features of cDNA Structure:
S4e is 1090 b p and has a 5' (55 bp) and a 3' (249 bp) noncoding region, along with a 786-bp open reading frame.
C/C Content:
41.4% (46.1 %O in coding region).
Structural Features of Proteins:
Open reading frame encodes 262 amino acids of which 58 are
basic. Predicted M, is 29,635. Putative bipartite nuclear targeting sequence from residues 239-256.
Antibodies:
None available.
using the program Clustal (Intelligenetics,Inc.). Cotton S4e
is 65,70,69, and 66% identical with S4e from yeast (Synetos
et al., 1992), mouse (Zinn et al., 1991), and human X and Y
chromosomes (Fisher et al., 19901, respectively. Also, we
have identified a putative bipartite nuclear-targeting sequence in cotton S4e. The presence of a nuclear-targeting
sequence is not surprising, since the biogenesis of ribosomal precursors (ribonucleoprotein particles) occurs
within the nucleus.
Ribosomal protein S4e is localized at the interface of the
40s and 60s subunits (Nygard and Nika, 1982; Uchiumi et
al., 1986), where it associates with initiation factor eIF-3
and may form part of the mRNA-binding domain (Westermann and Nygard, 1983). In plants, further work is
needed to characterize the genomic organization and regulation of S4e genes along with other genes encoding ribosomal proteins. Differences in ribosomal protein structure
(e.g. S4e) among plants, yeast, and animals could give us
Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does n o t imply its approval t o the exclusion o f other products that m a y also be suitable.
* Corresponding author; e-mail rtur1eyQasrr.arssuda.gov; fax
1- 601- 686 -9406.
431
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Turley e t al.
432
new insights into t h e importance of specific regions of
ribosomal proteins.
ACKNOWLEDGMENTS
The authors thank Shelia Meeks for her excellent technical assistance and Alex Kahler (Biogenetic Services) for the production
of restriction fragment length polymorphisms for cotton.
Received October 17, 1994; accepted November 14, 1994.
Copyright Clearance Center: 0032-0889/95/10S/0431/02.
The EMBL accession number for the sequence reported in this
article is X79300.
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Gao J, Kim S-R, Chung Y-Y, Lee JM, An G (1994) Developmental
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