GTPase Activity and Biochemical Characterization of a Recombinant Cotton Fiber Annexin1

A cDNA encoding annexin was isolated from a cotton (Gossypium hirsutum) fiber cDNA library. The cDNA was expressed in Escherichia coli, and the resultant recombinant protein was purified. We then investigated some biochemical properties of the recombinant annexin based on the current understanding of plant annexins. An “add-back experiment” was performed to study the effect of the recombinant annexin on β-glucan synthase activity, but no effect was found. However, it was found that the recombinant annexin could display ATPase/GTPase activities. The recombinant annexin showed much higher GTPase than ATPase activity. Mg2+ was essential for these activities, whereas a high concentration of Ca2+ was inhibitory. A photolabeling assay showed that this annexin could bind GTP more specifically than ATP. The GTP-binding site on the annexin was mapped into the carboxyl-terminal fourth repeat of annexin from the photolabeling experiment using domain-deletion mutants of this annexin. Northern-blot analysis showed that the annexin gene was highly expressed in the elongation stages of cotton fiber differentiation, suggesting a role of this annexin in cell elongation.

A Fiberless Seed Mutation in Cotton Is Associated with Lack of Fiber Cell Initiation in Ovule Epidermis and Alterations in Sucrose Synthase Expression and Carbon Partitioning in Developing Seeds1

Fiber cell initiation in the epidermal cells of cotton (Gossypium hirsutum L.) ovules represents a unique example of trichome development in higher plants. Little is known about the molecular and metabolic mechanisms controlling this process. Here we report a comparative analysis of a fiberless seed (fls) mutant (lacking fibers) and a normal (FLS) mutant to better understand the initial cytological events in fiber development and to analyze the metabolic changes that are associated with the loss of a major sink for sucrose during cellulose biosynthesis in the mutant seeds. On the day of anthesis (0 DAA), the mutant ovular epidermal cells lacked the typical bud-like projections that are seen in FLS ovules and are required for commitment to the fiber development pathway. Cell-specific gene expression analyses at 0 DAA showed that sucrose synthase (SuSy) RNA and protein were undetectable in fls ovules but were in abundant, steady-state levels in initiating fiber cells of the FLS ovules. Tissue-level analyses of developing seeds 15 to 35 DAA revealed an altered temporal pattern of SuSy expression in the mutant relative to the normal genotype. Whether the altered programming of SuSy expression is the cause or the result of the mutation is unknown. The developing seeds of the fls mutant have also shown several correlated changes that represent altered carbon partitioning in seed coats and cotyledons as compared with the FLS genotype.

Genes Involved in Osmoregulation during Turgor-Driven Cell Expansion of Developing Cotton Fibers Are Differentially Regulated1

Cotton (Gossypium hirsutum L.) fibers are single-celled trichomes that synchronously undergo a phase of rapid cell expansion, then a phase including secondary cell wall deposition, and finally maturation. To determine if there is coordinated regulation of gene expression during fiber expansion, we analyzed the expression of components involved in turgor regulation and a cytoskeletal protein by measuring levels of mRNA and protein accumulation and enzyme activity. Fragments of the genes for the plasma membrane proton-translocating ATPase, vacuole-ATPase, proton-translocating pyrophosphatase (PPase), phosphoenolpyruvate carboxylase, major intrinsic protein, and α-tubulin were amplified by polymerase chain reaction and used as probes in ribonuclease protection assays of RNA from a fiber developmental series, revealing two discrete patterns of mRNA accumulation. Transcripts of all but the PPase accumulated to highest levels during the period of peak expansion (+12–15 d postanthesis [dpa]), then declined with the onset of secondary cell wall synthesis. The PPase was constitutively expressed through fiber development. Activity of the two proton-translocating-ATPases peaked at +15 dpa, whereas PPase activity peaked at +20 dpa, suggesting that all are involved in the process of cell expansion but with varying roles. Patterns of protein accumulation and enzyme activity for some of the proteins examined suggest posttranslational regulation through fiber development.

Pacific Mills, Lawrence, Mass., Dover, N.H., Columbia, S.C... are the largest manufacturers in the world of cotton goods

Lawrence & Co.

Commonplace book, October 29, 1728-January 15, 1735

The small hardcover volume contains excerpts copied by John Winthrop beginning at the age of thirteen as a freshman at Harvard College. Winthrop’s own description of this book as “a book of Extracts from many Authors” is accurate, as it includes neither personal information about Winthrop nor reflections on what he read. With the exception of occasional notations indicating the date he finished reading particular books, the volume consists of passages he copied from a wide range of sources. It begins with an index of the various sections of the book (History, Poetry, Miscellanies, Geography, Divinity, History, Philosophy, Philosophy, Miscellanies, Poetry, Divinity, Philosophy) and ends with an index of the extracts' authors.

Payment order for Josiah Cotton, 1722 September 10

Handwritten order to Penn Townsend to pay scholarship funds to student Josiah Cotton (Harvard AB 1722), signed by Benjamin Wadsworth, Thomas Foxcroft, Samuel Marshall, and Jonathan Williams.

Payment order for Rowland Cotton, 1726 October 17

Handwritten order to Penn Townsend to pay scholarship funds to Rowland Cotton on behalf of his son Ward Cotton (Harvard AB 1729), signed by Thomas Foxcroft, John Marion, Samuel Marshall, and Jonathan Williams.

Ledger for the Classes of 1650-1663, 1649-1660

The leather-bound volume contains accounts for the Classes of 1650-1663 and Fellows. Many of the student bills were settled with payment in kind, often in the form of grain, apples, or livestock. The Steward's accounts with the College from 1656-1659 were entered on pages 295-303 and listed as "The steward is Creditor" and "The steward is Debitor." Credit entries include food, fuel, corn, and wages; debit entries list purchases and often identify the sellers.

Ledger for the Classes of 1732-1743, 1728-1749

The volume, bound in a modern green leather binding, contains accounts for the Classes of 1732-1743. The Steward's accounts with Harvard College are interspersed throughout the volume.

[Admittatur of Cotton Tufts (Harvard AB 1749)], 1745

Notebook with a handwritten copy of the 1734 College laws in English prepared by Harvard undergraduate Cotton Tufts and signed by President Edward Holyoke, Tutor Belcher Hancock, Fellow Joseph Mayhew, and Tutor Thomas Marsh on November 8, 1745.

Abstracts of sermons preached at Harvard College, 1728-1729

Small notebook containing notes kept by John Winthrop on sermons he attended between September 1, 1728 and October 19, 1729, while he was an undergraduate at Harvard College. The volume contains one-to-two page entries on specific sermons and provides the biblical text and related doctrines, questions, and conclusions. The inside back cover contains a handwritten index of the minister who gave the sermon, most often Nathaniel Appleton.

Mr. Monis's daggishes, [ca. 1728]

A one-page handwritten estimate of the "number & shape of ye letters wth dauguesh" needed to print the Hebrew Grammar sent by Judah Monis to the Corporation and a list of Hebrew characters with the related number of type needed for the printing. The document is undated but likely written in 1728 following the Harvard Corporation's vote on June 24, 1728 that the Treasurer should collect "so many Hebrew Types & points" needed for a complete set.