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BMC Mol Biol
2007 Mar 01;8:16. doi: 10.1186/1471-2199-8-16.
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Distinctive expression patterns of 185/333 genes in the purple sea urchin, Strongylocentrotus purpuratus: an unexpectedly diverse family of transcripts in response to LPS, beta-1,3-glucan, and dsRNA.
Terwilliger DP
,
Buckley KM
,
Brockton V
,
Ritter NJ
,
Smith LC
.
Abstract
BACKGROUND: A diverse set of transcripts called 185/333 is strongly expressed in sea urchins responding to immune challenge. Optimal alignments of full-length 185/333 cDNAs requires the insertion of large gaps that define 25 blocks of sequence called elements. The presence or absence of individual elements also defines a specific element pattern for each message. Individual sea urchins were challenged with pathogen associated molecular patterns (PAMPs) (lipopolysaccharide, beta-1,3-glucan, or double stranded RNA), and changes in the 185/333 message repertoire were followed over time.
RESULTS: Each animal expressed a diverse set of 185/333 messages prior to challenge and a 0.96 kb message was the predominant size after challenge. Sequence analysis of the cloned messages indicated that the major element pattern expressed in immunoquiescent sea urchins was either C1 or E2.1. In contrast, most animals responding to lipopolysaccharide, beta-1,3-glucan or injury, predominantly expressed messages of the E2 pattern. In addition to the major patterns, extensive element pattern diversity was observed among the different animals before and after challenge. Nucleotide sequence diversity of the transcripts increased in response to beta-1,3-glucan, double stranded RNA and injury, whereas diversity decreased in response to LPS.
CONCLUSION: These results illustrate that sea urchins appear to be able to differentiate among different PAMPs by inducing the transcription of different sets of 185/333 genes. Furthermore, animals may share a suite of 185/333 genes that are expressed in response to common pathogens, while also maintaining a large number of unique genes within the population.
Figure 1. Expression of 185/333 in response to various antigens. Nine animals were injected with either (A) LPS (n = 3), (B) Lam (n = 3), (C) dsRNA (n = 3), or (D) aCF (n = 2). Whole coelomic fluid (wCF) samples were collected prior to injection (pre) and at various times after challenge. The expression of both 185/333 (upper gels) and Sp056 (lower gels) were analyzed by RT-PCR in separate reactions and the agarose gels are shown. The vertical arrows indicate the time points at which the animals received injection(s). All animals received at least one injection at 0 h. Based on immune activation responses, three animals received second injections; animal 2-LPS and animal 5 at 24 h, animal 6 at 72 h.
Figure 2. Element patterns identified from sequences of the 185/333 amplicons. The element patterns (defined in [31]) for all of the clones isolated from this study are represented in graphical form. Different elements are shown as differently colored circles and ovals. Gaps are shown as horizontal lines. Short vertical black lines indicate the position of a stop codon and cDNAs with an early stop codon are designated by a decimal point followed by an integer. Elements with a diamond indicate the locations of indels that lead to a frame shift and missense amino acid sequence (elements with horizontal stripes). The sequences located 3' of the stop codon are shown to indicate that the remainder of the transcript sequence is present even though it may not be translated (vertical stripes). The frequency with which each pattern was identified (#) and the antigen that induced the pattern (Antigen) are indicated; L: LPS; β: Lam; R: dsRNA; a: aCF; *: whole bacterial challenge as identified in [31].
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